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Volume 19, Number 1 June 2000
SEP. 2] 2000

CURRENT HERPETOLOGY

FORMERLY THE JAPAMESE JOURNAL OF HERPETOLOGY

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4
Res
a

Jared.

Current Herpetology 19(1): 1-9., June 2000
© 2000 by The Herpetological Society of Japan

Predation by the Opossum Didelphis marsupialis on the

SELMA Maria ALMEIDA-SANTOS!, Marta Maria ANTONIAZZE,

! Laboratory of Herpetology, Instituto Butantan, Av. Vital Brazil 1500, 05503-900, Sdo
2 Laboratory of Cell Biology, Instituto Butantan, Ay. Vital Brazil 1500, 05503-900, Sado

3 Laboratory of Immunogenetics, Instituto Butantan, Av. Vital Brazil 1500, 05503-900,

Rattlesnake Crotalus durissus

OsvaLbo Aucusto SANTYANNA3, AND CARros JARED?*

Paulo, Brazil
Paulo, Brazil

Sado Paulo, Brazil

Abstract: Opossums are considered natural predators of snakes and possess
resistance to the venom of some viperids. The resistance of Didelphis to
Crotalus venom has been demonstrated through biochemical and immunologi-
cal assays. However, systematic observations on the behavior of adult Didel-
phis preying on venomous snakes have never been conducted. In this study
the predatory and defensive behaviors of Didelphis marsupialis and Crotalus
durissus, respectively, were analyzed in captivity. Defensive strategies showed
by snakes included immobility, flight attempts, coiling, cocking, rattling, and
counterattack with strikes and bites. The most common defensive behavior of
the rattlesnakes was immobility. The way the opossums attacked was clas-
sified in three categories, depending on the defensive reactions presented by the
snakes. On all occasions when the opossums were bitten, the injection of
venom apparently did not affect the predation. The great ability in capturing
and handling Crotalus durissus together with the apparent great tolerance to
the venom shown by Didelphis marsupialis when preying on these snakes
confirms the existent biochemical and immunological data about the resistance
of opossums to crotalic venoms. In this way our data strongly reinforce the
supposition that this species is an effective snake predator in nature.

Key words: Marsupialia; Behavior; Didelphis; Venom; Crotalus

INTRODUCTION (Cerqueira, 1985).

It is found in environ-

Didelphis marsupialis is a very common
marsupial in Brazil, living mainly in forests

* Corresponding author. Tel/Fax: +55 11
257-8905.

E-mail address: carlosjared@uol.com.br
Jared)

(C.

ments modified by humans, adapts well in
urban areas, and is nocturnal, scansorial,
and omnivorous. Its diet is composed of
fruits, nectar, small vertebrates, and inver-
tebrates (Emmons, 1990). Many reports
cite opossums as natural predators of
snakes including venomous species (Silva
Jr., 1956; Fitch, 1960; Cordero and Nicolas,

1986; Sazima, 1992). In addition, Brodie
III (1993) observed attacks of opossums
towards snake replicas in the field. At the
park of Butantan Institute, Sao Paulo,
Brazil, attacks by native opossums on the
outdoor enclosure snakes have been often
reported (W. Fernandes, pers. commun.).

A number of studies is available demon-
strating that opossums are immune to the
venom of some viperids (Domont et al.,
1991). Besides the well-known immunity
against Bothrops venom (Moussatché et al.,
1979, 1990; Perales et al., 1994; Neves-Fer-
reira et al., 1997), resistance of Didelphis to
the venoms of Crotalus durissus (Vellard,
1945; Moussatché et al., 1979, 1990) and C.
atrox (Werner and Vick, 1977; Perez et al.,
1979) has been observed. All these data
were obtained from biochemical and im-
munological assays. Studies about the zoo-
logical implications of this resistance have
never been undertaken.

Information about the predatory behav-
ior of American marsupials is scarce. Sazi-
ma (1992) made a single observation on the
behavior of Didelphis toward its prey; his
work, however, was not specific to preda-
tion. Jared et al. (1998) made a few prelimi-
nary observations on young opossums at-
tacking and killing young Bothrops jararaca
in captivity. Despite the strong supposition
about opossums being effective predators of
viperids, experimental results demonstrat-
ing this fact do not yet exist.

On the other hand, although snakes
present the most elaborate antipredator
mechanisms hitherto described among rep-
tiles, few papers are found about the defen-
sive behavior of these animals toward their
effective predators (Greene, 1973, 1988).
Among venomous snakes, rattlesnakes area
differentiated group possessing a unique
structure, the rattle, specialized in sonorous
defensive signaling (Greene, 1988). Ac-
cording to Duvall et al. (1985) and Greene
(1988), in nature the whole set of defensive
behaviors in rattlesnakes corresponds to an
increase of aggressiveness comprising

Current Herpetol. 19(1) 2000

procrypsis (immobility associated with a
cryptic coloration pattern), flight, body
coiling, cocking (retracting of the coiled
body and intimidation with strikes), rat-
tling, head hiding, strikes and bites, and
finally emptying of the anal glands. This
set of behaviors or, in most cases, the com-
bination of some of them, is sufficient to
prevent the snakes from being killed by pre-
dators.

This paper describes a behavioral experi-
ment in the laboratory where Crotalus
durissus was Offered to Didelphis marsupia-
lis. It aims to understand the defensive and
attack strategies of both animals. It also
has the intention of comparing the obtained
behavioral data with the existent biochemi-
cal and immunological information about
the resistance of Didelphis to Crotalus ven-
om. Drawing on these results, this work
finally tries to make a few inferences about
the predatory and defensive behaviors of
these animals in nature.

MATERIALS AND METHODS

Animals

Healthy adults D. marsupialis (N = 12)
collected from the woods of Butantan In-
stitute and from the surroundings of Sao
Paulo city were used. They weighed 1.5 to
2 kg.

The opossums were maintained in in-
dividual tanks measuring 0.80 ※ 0.70m
and 0.70 m in depth, and closed with a wire
netting lid. A wooden box measuring
0.27m in width, 0.34m in length, and
0.20 m in height with an inclined wire net-
ting front door was placed inside the tank
for shelter. Due to its design, the box did
not allow the opossum in the shelter to see
the environment inside the tank. This
shelter box occupied 16% of the total area
of the tank and was removed daily from the
tanks for cleaning, with the animal inside.
After that, the tanks were lined with
cardboard, and the box was replaced and
reopened. Water and food (fruits and

ALMEIDA-SANTOS ET AL.—OPOSSUM PREDATION ON RATTLESNAKES 3

minced meat) were supplied daily. Open-
ing and closing of the box door were done
with a hook-tipped stick.

Farmers from the State of Sao Paulo
(Brazil) supplied the snakes to the Herpetol-
ogy Laboratory of Instituto Butantan.
Thirty-six healthy adult Crotalus durissus
(total length = 60 to 90cm) were used.
They were maintained in captivity for at
least 20 days without being fed before the
experiments in order to guarantee that the
venom glands were full of secretion.

Behavioral evaluation

The observations were conducted during
a two-year period. Before initiating the ex-
periments each animal was kept in captivity
at least for a month for acclimation to the
new environment. Each opossum was test-
ed with one snake every 15 days for three
times per opossum. Before each observa-
tion the opossums were fasted for two days.
The experiments were conducted in dark-
ness at night, the period when the animals
are active. The tank was lighted inside with
a 15 W lamp and closed with a glass plate
instead of the netting lid. With this system
the opossums were kept acoustically and
visually isolated from the outside environ-
ment. At the same time, the inclined door
of the box made it possible for the observers
to see the opossum inside the box from top,
through the glass plate. The animal was
kept in this condition for 1 hr before the
snake was offered. The system allowed the
option for the opossum of attacking the
snake or remaining inside the box. Obser-
vations were started as soon as a snake was
gently placed into the tank. Twenty-five
out of a total of 36 experiments were
recorded on video tape and photographed.
In the experiments where predation did not
occur the total time of observation was 120
minutes. Five control experiments were
conducted placing a snake alone into the
tank and recording its behavior on video
tape for 1 hr.

A binomial test was used to statistically

test the preference for tail attack. The null
hypothesis was p = 0.5.

RESULTS

The opossum, even at night, constantly
remained inside the shelter box. When
food was placed in the tank, the opossum
immediately started to smell it with visible
movements of its snout and, in most cases,
came out of the box to feed on it. Some-
times, however, after smelling the food for
a few minutes it remained inside the box.

Two distinct phases of behavioral interac-
tions between the opossum and the rattles-
nake were characterized after the snake was
placed in the tank: 1) behaviors before the
attack, including the observations before
the opossum left the box and 2) the attack
itself, that was defined here as the set of be-
haviors observed from the moment the
opossum came out of the box and ap-
proached the snake until effective predation
occurred.

Before the attack, the opossum, inside
the box, immediately smelled the snake for
a few seconds to 2 min. During this time
different reactions of the snake could be ob-
served: 1) calm movements around the tank,
2) a quiet coiled posture, 3) immobility
(freezing), 4) flight, 5) threatening coiled
posture (cocking) or 6) rattling. The opos-
sum, after smelling the snake, sometimes
remained in the box, not leaving to catch it.
In four experiments, when the opossum in-
side the box spent more time smelling the
snake, rattling was observed; in all these oc-
casions the opossum chose to remain in the
box, not attacking the snake.

When the opossum attacked, it came out
of the box, approached the snake quickly
(Fig. 1A), and captured it (Fig. 1B). On
three occasions, the attack was so quick that
the snake had no time to react. Most times,
however, the snake showed some type of
reaction.

Three types of behavioral interactions
were recognized during the attacks. Figure

4 Current Herpetol. 19(1) 2000

Fic. 1. One of the possible sequences of predatory behaviors of the opossum on the rattlesnake.
A: The opossum directly approaches the snake, which remains immobile. B: The opossum captures
the snake by the tail. The snake remains passive, flicking the tongue. C: The opossum continues
eating the snake from the tail. The snake still flicks its tongue. D: The snake suddenly bites the
opossum. E: The opossum reacts, kills the snake by chewing its head (arrow) and bites the rest of the
body. F to H: The opossum continues eating the snake from the anterior end. While eating, it
remains in the same position (F, G) until it finishes the whole snake (H).

ALMEIDA-SANTOS ET AL.—OPOSSUM PREDATION ON RATTLESNAKES 5

2 summarizes the feeding tactics of the
opossums, which depended on the reactions
exhibited by the snakes after the initial ap-
proach:

(1) The snakes showed an erratic behav-
ior, moving the body by chance in a
disoriented manner. In this case, the
opossum immobilized them with suc-
cessive bites along the whole body.
Then it consumed them from either the
tail or the head.

The snakes were immobile. In this
case the opossum consumed them
alive, preferentially from the tail (Fig.
1C). Usually the snakes stayed immo-

(2)

bile throughout predation. Some-
times, however, the snakes counterat-
tacked with part of their bodies al-
ready eaten.
The snakes counterattacked with
strikes and bites either just before
being captured or having already part
of its body eaten from the tail (Fig.
1D). In both cases the opossum killed
them by chewing the head, and then
immediately consumed them beginning
from one of the extremities (Fig. 1E).
Besides these three types of interactions, a
few encounters were observed where the
opossum and the snake faced each other

(3)

THE OPOSSUM RECOGNIZES THE PRESENCE OF THE
SNAKE THROUGH SMELL

N=25

THE OPOSSUM INCREASES
SMELLING AND ATTACKS

1. THE SNAKE
EXHIBITS ERRATIC
BEHAVIOR

THE OPOSSUM
IMMOBILIZES THE
SNAKE THROUGH

SEVERAL BITES ALONG

THE BODY AND BEGINS
EATING FROM THE

HEAD OR FROM THE

TAIL

2. THE SNAKE
STAYS IMMOBILE

THE OPOSSUM EATS
THE SNAKE ALIVE,
PREFERABLY BEGINNING
FROM THE TAIL

THE SNAKE
REMAINS
IMMOBILE

THROUGHOUT
PREDATION

N=9

3. THE SNAKE
COUNTERATTACKS
BY STRIKING

N=12

N=11

THE OPOSSUM KILLS
THE SNAKE BY CHEWING
THE HEAD AND
BEGINS EATING FROM
THE HEAD OR
FROM THE TAIL

Fic. 2. Scheme of attack and defense strategies in the encounters of opossums and snakes. Three
different types of reaction are observed in the rattlesnakes after the first attack by the opossum. N =

number of filmed events.

without moving for a long time. The two
animals remained in this position until the
snake slowly moved backwards.

During predation, the prey handling be-
havior was quite regular: the opossum
remained in a sitting position while holding
the snake with one of the forelimbs (Figs.
1F, 1G). One of the ends of the snake was
introduced into the mouth laterally and was
chewed with the lateral teeth (Figs. 1F, 1G).
The other forelimb supported the animal
(Figs. 1F, 1G). The forefeet were used al-
ternately but for short periods of time were
used together to tear apart the harder parts
of the snake. Generally the whole snake
was eaten uninterruptedly (Fig. 1H) with an
average duration of 12 min.

The opossums killed and ate approxi-
mately 80% of the offered rattlesnakes.
Taking into account the filmed captures
where the attack was directed to the tail or
to the head (N = 22), the opossums seemed
to direct the attack significantly to the tail
(N = 16; p < 0.05, df = 1). In three
filmed captures the opossums directed the
attack to the middle of the body.

During all experiments no observable
effects of envenomation were recognized.

DISCUSSION

The way the opossums behaved while eat-
ing the snakes was very regular. The opos-
sums were always in the same posture when
observed. They used only one forelimb at a
time to hold the snake and introduce it into
the mouth. Ivanco et al. (1996) also
verified the use of a single limb in another
opossum species, Monodelphis domestica,
when feeding or preying, and suggested that
this behavior is fixed and species-typical.

During the time preceding the effective
predation, when the encounter between
opossum and rattlesnake took place, great
behavioral variations were noted in both
animals.

Before the attack by the opossum, the
differences in reaction presented by the rat-

Current Herpetol. 19(1) 2000

tlesnakes indicated that some times they
were not able to notice the opossum, mov-
ing calmly around the tank or remaining
quietly coiled. Most times, however, the
rattlesnakes demonstrated by their behavior
that they could recognize the opossum as a
predator; in these cases some of the typical
behaviors of the defensive escalation of the
rattlesnakes were observed, such as
immobility, coiling, cocking, and rattling.
Among these defensive behaviors, the
strategy of immobility in nature can be very
valuable when associated with a cryptic
coloration pattern and may constitute an
efficient defense used by these animals
against predators (Greene et al., 1978; Her-
zog and Drummond, 1984; Cloudsley-
Thompson, 1994). Rattling many times oc-
curred despite the snake not being able to
see the opossum inside the box. Since it
was not observed in any of the control ex-
periments, rattling indicates that most times
the snakes seemed to identify somehow the
danger, possibly by chemical signs, as
proposed by Weldon et al. (1992).

On a significant number of occasions the
attack of the opossum was directed to the
tail. That was observed more frequently
when the rattlesnakes remained immobile, a
strategy that at first view is difficult to inter-
pret. On the other hand, during the ap-
proach, it was observed on a few occasions
that the opossum, when noticing the snake
coiling and preparing to strike, rapidly
killed it by attacking and chewing its head.
The same happened in the few cases when
the rattlesnakes were able to bite the opos-
sum even after being captured by the tail.
In other cases, when the snake reacted to
the attack by exhibiting erratic behavior, it
was immobilized through bites along the
whole body. The analysis of these different
situations observed during the attack indi-
cated that any type of active reaction
presented by the snake caused an immediate
fatal attack by the opossum. Although in
our experimental conditions attacks direct-
ed to the tail did not prevent the snake from

ALMEIDA-SANTOS ET AL.—OPOSSUM PREDATION ON RATTLESNAKES 7

being killed by the opossum, in nature they
may confer an advantage to the snake, that
by remaining immobile has a chance of es-
caping without being severely injured, as
has been already observed for lizards by
Greene et al. (1978). These data seem to be
in accordance with Herzog and Burghardt
(1974), who affirmed that for many preda-
tors, prey movement is a critical factor in
mediating attack.

In contrast to our results indicating some
preference of the opossums for capturing
the snakes by the tail, Sazima (1992) report-
ed that D. marsupialis when attacking
Bothrops jararaca usually goes first to the
head or neck region. In our experiments
with Crotalus durissus, in many cases,
Didelphis grasped the tail first, giving the
snakes a chance to bite. This observation,
at first view, seems to be contradictory since
the predatory behavior of ophiophagous
animals (mammals or birds) usually consists
in attacking the head or the region just be-
hind the head (Kaufmann and Kaufmann,
1965; Perez et al., 1978). However,
ophiophagous animals such as Conepatus
sp. and Galicts sp. have been observed at-
tacking snakes at the tail (Ribeiro, 1940;
Jackson, 1979).

It is possible that in our study the appar-
ent preference of D. marsupialis for attack-
ing the tail of C. durissus may be caused by
an attraction of the opossums to cloacal
odors of the rattlesnakes that can misdirect
the attack. In nature, such attraction of
the predator to the tail, which is a more dis-
posable portion of the body, could help the
prey to escape or counterattack (Greene,
1988; Alcock, 1993).

Two species of opossums of the genus
Didelphis occur in Brazil: D. albiventris and
D. marsupialis (Cerqueira, 1985). The
former lives in open fields such as “cerrado”
and “caatinga” and the latter is distributed
in forests (Cerqueira, 1985; Emmons,
1990). On the other hand, Crotalus duris-
SUS 1S a Species typical of open fields while
Bothrops jararaca is distributed in forests

(Sazima, 1992; Campbell and Lamar, 1989).
In this way, one would expect the opossums
to have resistance only to snake venoms
from the same habitat. In fact, Mous-
satché et al. (1990) have demonstrated that
D. marsupialis remains unharmed by B.
jararaca venom. In addition, they men-
tioned that this marsupial has partial
resistance to Crotalus durissus venom. On
the basis of this information, our experi-
ments aimed at comparing behavioral
results with the biochemical data of Mous-
satché et al. (1990). Although D. marsu-
pialis and C. durissus are not sympatric in
nature, we observed that, at least in captivi-
ty, predation was effective. The injection
of venom by snakebites apparently did not
affect the predation as has already been ob-
served for other predators, including mam-
mals and birds (Duvall et al., 1985).

In spite of the limitations imposed by a
behavioral experiment conducted in captivi-
ty, the present observations strongly suggest
that D. marsupialis is an effective snake
predator in nature. This supposition is
mainly based on the great interest and abili-
ty shown by D. marsupialis in capturing
Crotalus durissus, which were comparable
to the interest they showed when presented
with different types of food. This idea is
also reinforced by the great tolerance these
animals showed to the snake venom.

ACKNOWLEDGEMENTS

The authors wish to thank Ms. Simone
Jared, Mrs. Laurinda A. Soares, Mrs.
Alaide M. Marques, Mr. Valdir J. Germano
and Mr. Yutaka Kanasi, for their valuable
technical assistance. We are also thankful
to Dr. André Eterovic and Mr. Lucio
Parana Catani for suggestions. Fundacao
Butantan and FAPESP (proc. 98/8183-5)
provided the financial support. Dr. Os-
valdo Augusto Sant’Anna is supported, in
part, by CNPq Brazil.

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of the opossum to some snake venoms. Toxi-
con 17: 130.

MoUssATCHE, H., J. PERALES, A. G. C. N. FER-
REIRA, S.L.G. RocuHa, C.G. VILLELA, AND
G.B. Domont. 1990. Resisténcia de mar-
supiais brasileiros aos venenos de Bothrops
jararaca e Crotralus durissus terrificus. Proc.
Vth Ann. Meet. Fed. Soc. Exp. Biol., MG,
Brazil. p. 397.

NEVES-FERREIRA, A. G., J. PERALES, M. Ova-
DIA, H. MOUSSATCHE, AND G.B. DOMONT.
1997. Inhibitory properties of the an-
tibothropic complex from the South American
opossum (Didelphis marsupialis) serum. Toxi-
con 35: 849-863.

PERALES, J., H. MOUSSATCHE, S. MARANGONI,
B. OLIVEIRA, AND G. B. DoMoNr. 1994. Iso-
lation and partial characterization of an anti-
bothropic complex from serum of South
American Didelphidae. Toxicon 32: 1237-
1249.

PEREZ, J.C., H.C. Haws, V.E. Garcia, AND
B. M. JENNINGS, III. 1978. Resistance of
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PEREZ, J. C., S. PICHYANGKUL, AND V. E. GAR-
cIA. 1979. The resistance of three species of
warm-blooded animals to western diamond-
back rattlesnake (Crotalus atrox) venom. Tox-
icon 17: 601-607.

RIBEIRO, L. 1940. Medicina no Brasil. Imprensa
Nacional, Rio de Janeiro. 409 p.

ALMEIDA-SANTOS ET AL.—OPOSSUM PREDATION ON RATTLESNAKES 9

SAZIMA, I. 1992. Natural history of the jararaca
pitviper, B. jararaca in Souheastern Brazil.
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Brodie, Jr. (eds.), Biology of the Pitvipers.
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SrLVA,。 M. Jr. 1956. O Ofidismo no Brasil.
Ministério da Saude, Rio de Janeiro. 346 p.
VELLARD, J. 1945. Resistencia de los “Didel-
phis” (Zarigueya) a los venenos ofidicos. Rev.

Bras. Biol. 5: 463-467.
WELDON, P.J., R. ORTIZ, AND T.R. SHARP.

1992. The Chemical Ecology of Crotalinae
Snakes. p. 309-319. In: J. A. Campbell and
E. D. Brodie, Jr. (eds.), Biology of the Pit-
vipers. Selva Publ., Tyler, Texas.

WERNER, R.M. AND J.A. Vick. 1977.
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Accepted: 23 December 1999

の oa 5
還 本 2PAM2HTTTA VO MOTTACB2G UGCGOIRES

と a

Current Herpetology 19(1): 11-14., June 2000
© 2000 by The Herpetological Society of Japan

Aphaniotis nasuta (de Jong, 1930), a Junior Synonym
of A. ornata (Van Lidth de Jeude, 1893)
(Squamata: Agamidae)

HIpDETOSHI OTA!* AND Tsutomu HIKIDA2

1 Tropical Biosphere Research Center, University of the Ryukyus, Nishihara, Okinawa,
903-0213 JAPAN
2 Department of Zoology, Graduate School of Science, Kyoto University, Kitashiraka-
wa, Sakyo, Kyoto, 606-8502 JAPAN

Abstract:

Morphological comparisons of available types of the two snout-

ornamented agamids from northern Borneo, Aphaniotis ornata and A. nasuta,
failed to show any substantial differences. Careful examination of original
descriptions of these nominate taxa also yielded no discriminant characters.
Thus, although the holotype of A. nasuta was not detected in our survey of
various museum collections, we are sure that it is appropriate to synonymize
this nominate species with A. ornata.

Key words:
Borneo

INTRODUCTION

Van Lidth de Jeude (1893) described
Japalura ornata on the basis of a female
agamid with long limbs and a rostral ap-
pendage from near Sandakan Bay, North
Borneo (i.e., Sabah, Malaysia). He as-
signed this species to Japalura chiefly be-
cause of the presence of an oblique fold in
front of the shoulder. Later, de Jong
(1930) described another long-limbed,
snout-ornamented agamid species, Japalura
nasuta, on the basis of six specimens also
from North Borneo. He, however, did not
compare this species with J. ornata, nor

* Corresponding author. Tel: +81-98-895-
8937; Fax: +81-98-895-8966.

E-mail address: ota@sci.u-ryukyu.ac.jp. (H.
Ota)

Aphaniotis nasuta; Aphaniotis ornata; Agamidae; Synonymy;

even mention the latter.

In his doctoral dissertation, Moody
(1980) made drastic changes in the clas-
sification of the family Agamidae, which in-
volved translocations of ornata and nasuta
from Japalura to another genus, Aphanio-
tis. He did not mention any concrete rea-
son for such rearrangements, but it is
almost certain that these and other changes
proposed in his taxonomic list (given as Ap-
pendix A) reflect his morphological re-
definitions of genera resulting from
phylogenetic analyses of the whole family
(Moody, 1980).

Despite the ambiguity regarding the
validity of nasuta in the presence of ornata
(see above), almost no one subsequent to de
Jong (1930) has addressed this taxonomic
problem, and only Manthey and Gross-
mann (1997) pointed out the necessity for

the clarification of their differences. Thus,
both ornata and nasuta have been literally
regarded as valid species of Japalura (e.g.,
Wermuth, 1967), or of Aphaniotis (e.g.,
Welch et al., 1990; Welch, 1994).

Recently we examined type specimens of
both of these species. The Results of com-
parisons strongly suggest that these
nominate species are actually conspecific.

MATERIALS AND METHODS

The holotype of A. ornata (RMNH 4344:

Current Herpetol. 19(1) 2000

Fig. la) and a paratype of A. nasuta (SMF
78702: Fig. 1b), both adult females, were
examined. Definitions of quantitative
characters follow Ota (1991). Institutional
acronyms are those suggested by Leviton et
al. (1985).

RESULTS AND DISCUSSION

Table 1 compares 15 quantitative charac-
ters of the types of A. ornata and A. nasuta.
These specimens greatly resembled each
other in most characters examined. The

Fic. 1.
nasuta (SMF 78702, SVL = 53.5 mm) (b).

Holotype of Aphaniotis ornata (RMNH 4344, SVL = 54.2 mm) (a), and a paratype of A.

OTA & HIKIDA—AGAMID LILARD SYNONYMY 13

TABLE 1. Comparisons of meristic and morphometric characters (in mm) between holotype of
Aphaniotis ornata (RMNH 4344) and a paratype of A. nasuta (SMF 78702). Abbreviations are as fol-
lows. SL: supralabials; IL: infralabials; IO: interorbital scales; MSR: scale rows around midbody;
FIVS: finger IV subdigital scales; TIVS: toe IV subdigital scales; SVL: snout-vent length; HL: head
length; HW: head width; SFL: snout-forelimb length; AGL: axilla-groin length; FLL: forelimb length;
HLL: hind-limb length; FIVL: finger IV length; TIVL: toe IV length.

Character A. ornata A. nasuta
SL 8 8

IL vi

IO 19 Dips

MSR 80 7 が /

FIVS 18 19

TIVS 2 24

SVL 54.2 5325

HL (ratio to SVL) 14.0 (25.8%) en OLA,

HW (ratio to SVL)
SFL (ratio to SVL)
AGL (ratio to SVL)
FLL (ratio to SVL)
HLL (ratio to SVL)
FIVL (ratio to SVL)
TIVL (ratio to SVL)

9.9 (18.3%)
22.4 (41.3%)
25.5 (47.0%)
29.0 (53.5%)
51.1 (94.3%)

7.6 (14.0%)
10.6 (19.6%)

9.7 (18.1%)
21.1 (39.4%)
24.4 (45.6%)
31.0 (57.9%)
54.9 (102.6%)

7.8 (14.6%)
5 人 (2690

possible greatest differences lay in the rela-
tive fore- (FLL) and hindlimb lengths
(HLL) that were somewhat greater in the
paratype of A. nasuta (57.9% and 102.6%
of the snout-vent length [SVL], respective-
ly) than in the holotype of A. ornata (53.5%
and 94.3%, respectively). Even so, such
differences, corresponding to 4.4% of SVL
in FLL and 8.3% of SVL in HLL, are well
within the extent of variations in cor-
responding characters among conspecific fe-
males from limited geographical ranges
reported for other arboreal agamids (e.g.,
5.9% of SVL in FLL and 9.3% of SVL in
HLL for Calotes cristatellus from Sabah
[Ota and Hikida, 1991], and 8.5% of SVL
in FLL and 12.7% of SVL in HLL for
Japalura swinhonis from Taiwan [Ota,
1991]).

Between the two specimens, there were
also no differences evident in qualitative
characters, such as the shape of the rostral
appendage and overall coloration. Careful

comparisons of original descriptions of
Aphaniotes ornata and A. nasuta (e.g., Van
Lidth de Jeude [1893] and de Jong [1930],
both as Japalura: see above) also failed to
reveal any substantial differences between
these nominate species.

According to de Jong (1930), the type
series of A. nasuta consisted of two males
(including the holotype) and four females,
all deposited in the Buitenzorg Museum,
Java. This museum was largely succeeded
by Museum Zoologicum Bogoriense (MZB)
after World War II. Indeed, the SMF
specimen examined by us was labeled as
“Formerly in Mus. Bogor.” However,
despite our intensive survey of various
museum collections including that of MZB,
we did not find the holotype or any of the
remaining paratypes of A. nasuta.

We consider it to be best at present to
synonymize A. nasuta with A. ornata, be-
cause comparisons both of available types
and of original descriptions strongly suggest

Current Herpetol. 19(1) 2000

their identity as mentioned above. Further
efforts should be made to detect other types,
especially the holotype, of A. nasuta to
verify this account.

ACKNOWLEDGMENTS

We thank M. S. Hoogmoed (RMNH) and
G. Kohler (SMF) for the arrangement to ex-
amine specimens under their care. H. Ota
also thanks the staff of MZB for responding
to his query regarding the types of Aphani-
otis nasuta. This research was supported
by a Grant-in-Aid from the Japan Ministry
of Education, Science, Sports and Culture
(Oversea Research No. 10041166) through
the courtesy of M. Matsui.

LITERATURE CITED

DE JONG, J.K. 1930. Notes on some reptiles
from the Dutch-East-Indies. Treubia 12(1):
115-119.

DE Roo, N. 1915. The Reptiles of the Indo-
Australian Archipelago. I. Lacertilia, Chelo-
nia, Emydosauria. E. J. Brill, Leiden. 384 p.

LEvITON, A. E., R. H. Gress, Jr., E. HEAL, AND
C. E. Dawson. 1985. Standards in herpetolo-
gy and ichthyology: Part I. Standard symbolic
codes for institutional resource collections in
herpetology and ichthyology. Copeia 1985(3):
802-832.

MANTHEY, U. AND W. GROSSMANN. 1997. Am-
phibien & Reptilien Sudostasiens. Natur und
Tier - Verlag, Munster, Germany. 512 p.

Moopy, S. M. 1980. Phylogenetic and Histori-
cal Biogeographical Relationships of the
Genera in the Family Agamidae (Reptilia:
Lacertilia). Unpublished Ph.D. Dissertation.
University of Michigan, Ann Arbor. 373 p.

Ota, H. 1991. Taxonomic redefinition of
Japalura swinhonis Gunther (Agamidae:
Squamata), with a description of a new sub-
species of J. polygonata from Taiwan. Her-
petologica 47(3): 280-294.

Ota, H. AND T. HrIkrpA. 1991. Taxonomic re-
view of the lizards of the genus Calotes Cuvier
1817 (Agamidae Squamata) from Sabah,
Malaysia. Trop. Zool. 4(2): 179-192.

VAN LIDTH DE JEUDE, T. W. 1893. On reptiles
from North Borneo. Notes Leiden Mus. 15:
250-257.

WELCH, K. R. G. 1994. Lizards of the World: A
Checklist. 5. Agamidae, Chamaeleonidae,
Cordylidae and Gerrhosauridae. KCM Books,
Somerset. 97 p.

WELcH, K.R.G., P.S. CooKE, AND A.S.
WriGHT. 1990. Lizards of the Orient: A
Checklist. R.E. Krieger Publ., Malabar,
Florida. 162 p.

WERMUTH, H. 1967. Liste der rezenten Am-
phibien und Reptilien. Agamidae. Das Tier-
reich 86: 1-105.

Accepted: 26 January 2000

Current Herpetology 19(1): 15—26., June 2000
© 2000 by The Herpetological Society of Japan

Conspecific and Heterospecific Pair-formation in Rana

porosa brevipoda and Rana nigromaculata, with
Reference to Asymmetric Hybridization

RYOHEI SHIMOYAMA

Laboratory of Animal Ecology, Joetsu University of Education, 1 Yamayashiki, Joetsu,
Niigata 943 JAPAN

(Present address: 2599-6 Yonezawa, Chino, Nagano 391-0216 JAPAN)

Abstract:

Patterns of conspecific and heterospecific pair-formation in Rana

porosa brevipoda and Rana nigromaculata were investigated in the field. In R.
porosa brevipoda, most of the conspecific pairs were formed by female initia-
tion. Size-assortative mating was observed in R. porosa brevipoda. On the
other hand, most of the conspecific pairs of R. nigromaculata were formed by
forced clasping by males. There was no significant correlation between the
male and female sizes in amplectant pairs of R. nigromaculata. Of a total of
12 heterospecific pairs observed, 11 were pairs between male R. nigromaculata
and female R. porosa brevipoda. The proximate factors of the asymmetry in
heterospecific pairing are discussed with relation to the differences in the pair-
ing patterns, body size, and the body shape between the two species. Possible
impacts of the asymmetric hybridization on the two species are also discussed.

Key words: Rana porosa brevipoda; Rana nigromaculata; Pair-formation;
Heterospecific pairing; Hybridization

INTRODUCTION

The Japanese pond frogs Rana porosa
brevipoda and Rana nigromaculata are
closely related phylogenetically and have
similar ecological requirements (Maeda and
Matsui, 1989). Postmating reproductive
isolating mechanisms between these two
species are quite incomplete. Although
male hybrids are sterile, females hybrids are
mostly fertile (see Maeda and Matsui, 1989;

Tel: 十 81-266-73-7766.
E-mail address: rshirmo(②pob.lcv.ne.jp

Matsui, 1996 for review). Currently,
natural hybridization between these two
species has been reported at numerous
localities of their sympatric ranges (Nishio-
ka et al., 1981, 1992). However, there is no
study on the ethological and ecological fac-
tors causing natural hybridization between
these two species.

In the northern part of the Ina Basin,
Nagano Prefecture (central Japan), an iso-
lated population of R. porosa brevipoda is
distributed sympatrically with R. nigro-
maculata, which has a wide range of distri-
bution (Nishioka et al., 1981; Shimoyama,

1986a). In this region, frogs with inter-
mediate external characters, which are con-
sidered to be natural hybrids, are also found
(Shimoyama, 1986a, 1996). Actually,
Nishioka et al. (1981, 1992) showed evi-
dence of introgressive hybridization be-
tween the two species in this region based on
the electrophoretic analyses of allozymes
and blood proteins.

In the studies of comparative reproduc-
tive ecology and interspecific relationships
of the two pond frogs, Shimoyama (1996)
clarified the absence of premating reproduc-
tive isolation between the two species in the
northern Ina Basin. Shimoyama (1996)
pointed out the absence of distinct segrega-
tion in diel, seasonal, and spatial patterns of
breeding activity between the two species,
which results in the formation of mixed-
species choruses comprising males of both
species. Such a mixed-species chorusing is
caused by the misidentification of hetero-
specifics as conspecifics by male R. nigro-
maculata (Shimoyama, 1999). Shimoyama
(1999) further documented similarities of
vocal repertoires, advertisement call struc-
tures, and male social behavior between the
two species. In addition, Shimoyama
(1999) demonstrated the occurrence of het-
erospecific pairing and spawning within the
mixed-species choruses.

In the present paper, I first describe pat-
terns of conspecific and heterospecific pair-
formation of the two species, with relation
to the asymmetry in the heterospecific pairs
and hybridization. I then discuss the possi-
ble proximate factors and influences of the
asymmetric hybridization.

METHODS

Study area

This study was done in the breeding sea-
sons of 1990, 1992, 1993, and 1995-1997 in
Tatsuno (35°56 N, 137°58 E, 720m above
sea level), Nagano Prefecture, central
Japan. A detailed description of the study
site is given in Shimoyama (1996).

Current Herpetol. 19(1) 2000

Observations

Field survey was made on 47 days in
May-July 1990, five days in May 1992, six
days in May 1993, eight days in May-July
1995, 18 days in May-June 1996, and six
days in May-June 1997. Details of the
general methods are described in Shimoya-
ma (1996, 1999).

Observations on male behavior within the
mixed-species choruses were made usually
at 0500-0700 hr, when males and females of
both species are most active (Shimoyama,
1996). When breeding activities were still
very intense after O800hr, observations
were continued until 1000 hr. The sum of
behavioral observations was 265 hours. On
every visit, I selected one or two mixed-spe-
cies chorus(es) for observation of male
and/or female behavior from _ nearby
(usually < 5m) banks. Binoculars (x7)
were used to ascertain individual identifica-
tion. When gravid females were found
around or within the choruses, I started the
focal animal sampling (Martin and Bateson,
1986) for the females until they finished
spawning.

After spawning, the number of eggs wi-
thin each clutch was counted for the clutch-
es of R. porosa brevipoda to determine
whether it was a first or second clutch (see
Serizawa, 1983, Shimoyama, 1986b). I
could judge the clutches to be first or second
based on the relationship between female
snout-vent length (hereafter SVL) and the
number of eggs within the clutch (see below
and Serizawa et al., 1990).

When heterospecific spawnings were ob-
served, I monitored the egg masses for
several days to determine whether they
hatched normally or not. If at least a part
of the eggs hatched, I considered that
hybridization had occurred.

RESULTS AND DISCUSSION

Breeding of R. porosa brevipoda
Size distribution of mature R. porosa
brevipoda is shown in Fig. 1. Data from

SHIMOYAMA—FROG PAIR-FORMATION

40
に
で

っ 30
iS
=
a2)

に 20
oO

o 10
z

40 45 50 55 60 65 70 TK 80
SVL (mm)
Fic. 1. Size distribution of mature individuals of Rana porosa brevipoda. Open and closed

histograms show males and females, respectively.

the seven seasons were pooled. Histograms
of the mature males (hereafter males) show
two distinct size groups. One is composed
of smaller individuals with SVL of 40-
50 mm, and the other is composed of larger

SVL (mm)

individuals with SVL of more than 50 mm.
The former group is estimated to be com-
posed of 1-yr-olds, and the latter to be
composed of older animals (see Inoue,
1979; Serizawa, 1983; Shimoyama, 1989).

Days since 10 May

Fic. 2. Relationship between body size and the date males of R. porosa brevipoda were first found

calling in 1990.

On the other hand, mature females (here-
after females) were not divided into distinct
size groups. Females were estimated to be
older than 1 year (Shimoyama, 1986b,
1989). Mean SVL of male and female R.
porosa brevipoda was 55.7 mm (SE=0.31,

= 344) and 62.5 mm (SE=0.55, N=130),
respectively. Females were significantly
larger than males (Mann-Whitney U-test,
z=9.03, p=0.0001).

Figure 2 shows the relationship between
the date of first discovery of calling and the
male SVL. There was a significant tenden-
cy for smaller males to begin calling later in
the season than larger males (r= —0.332,
N=179, p=0.0001). As shown in Fig. 3,
there was also a significant trend for larger
males to call on more nights than smaller
males (r=0.364, N=179, p=0.0001).

On the basis of the relationship between
female SVL and number of eggs within the
clutch, I could judge whether each clutch

45
40
35
30
25
20

No. of nights called

Current Herpetol. 19(1) 2000

was a first or second one (Fig. 4; see also
Serizawa et al., 1990). The average number
of eggs within the estimated first clutches
and the second ones were _ 1813.0
(SE=61.42, N=28) and 556.8 (SE=57.56,
N=13), respectively. Figure 5 shows the
relationship between the date of oviposition
and the female SVL. There was a trend for
larger females to deposit first clutches earli-
er in the season and deposit second clutches
approximately a month later. But deposi-
tion of the second clutch was not observed
in females of less than 63 mm SVL.

During the course of the study, I found
62 conspecific pairs of R. porosa brevipoda.
These pairs included unmarked individuals,
whose size was not known. I observed the
sequence of the pair-formation for 36 cases.
As reported in Shimoyama (1993b), two
major patterns of the pair-formation were
found. One was female initiation, and the
other was forced clasping by males. Of the

。9 <

eo ep の

ーー こい

55 60 65

SVL (mm)

Fic. 3.
brevipoda in 1990.

Relationship between body size and the number of nights of calling in males of R. porosa

SHIMOYAMA—FROG PAIR-FORMATION 19

@ ist clutch
Oo 2nd clutch

2500

2000

1500

1000

Clutch size

500

50 55 60 65 70 75 80
SVL (mm)

Fic. 4. Relationship between female size and the number of eggs per clutch in R. porosa brevipo-
da. Data from the seven seasons were pooled. Regression equation for the estimated first clutch is:
Y=58.5X—1996.9 (r=0.941, N=28, p=0.0001), and that for the estimated second clutch is:
Y =35.4XK — 1880.7 (r=0.617, =13, p=0.0247).

SVL (mm)

50
0 10 20 30 40 50 60

Days since 10 May

Fic. 5. Relationship between the date of oviposition and the size of female R. porosa brevipoda on
1990. First clutch: r= —0.592, N=28, p=0.0009; second clutch: r=0.06, N=13, p>0.8.

36 cases of the pairing sequence, 30 (83.3%)
were initiated by the female. In all of the
30 cases, a gravid female visited only one
calling male and mated with the first male
she visited. Gravid females seemed to ap-
proach males which initiated bouts of call-
ing more frequently. The remaining six
cases of the pairing sequence were initiated
by forced clasping by males: four and two
females were clasped by calling males and
satellite males, respectively (see Shimoya-
ma, 1993b).

Figure 6 shows the relationship between
the SVL of females and that of males in the
conspecific pairs of R. porosa brevipoda.
There was a significant positive correlation
between the male and female sizes (r=
0.534, N=61, p=0.0001). This phenomen-
on, which is called “size-assortative mat-
ing’, has been considered to be evidence of
female mate choice. However, Arak (1983)
pointed out that size-assortative mating

SVL of males (mm)

50 95 60

Current Herpetol. 19(1) 2000

alone cannot be regarded as evidence of
mate choice by females, but that male-male
competition could incidentally make a pat-
tern of size-assortative mating. In the
present study, I did not obtain adequate
data to further discuss the proximate and
ultimate factors of the size-assortative mat-
ing observed in R. porosa brevipoda.

Breeding of R. nigromaculata

Figure 7 shows the size distribution of
mature R. nigromaculata. SVL of males
ranged from 49.0 to 80.8 mm with a mean
of 66.0mm (SE=0.39, N=172), and that
of females from 60.0 to 83.0mm with a
mean of 70.9mm (SE=0.67, N 三 62).
Males were estimated to be more than 1
year of age, and females to be more than 2
years of age (Shimoyama, 1989). Distinct
size groups were not found in either sex.
Females were significantly larger than
males (Mann-Whitney U-test, z=5.86, p=

65 70 198 80

SVL of females (mm)

Fic. 6. Relationship between the male and female sizes in amplectant pairs of R. porosa brevipo-

da. Data from seven seasons were pooled.

SHIMOYAMA—FROG PAIR-FORMATION 21

No. of individuals

45 50 55 60 65 70 TES) 80 85
SVL (mm)

Fic. 7. Size distribution of mature individuals of Rana nigromaculata. Open and closed histo-
grams show males and females, respectively.

0.0001). cy for larger males to call on more nights

There was no trend between the date of than smaller males (r=0.598, N=84,
the first discovery of calling and male size in p=0.0001; Fig. 9). Figure 10 shows the
R. nigromaculata (r=0.048, N= 84, p > 0.6; relationship between the date of oviposition
Fig. 8). But there was a significant tenden- and female SVL in 1990. There was a sig-

eee る ぐ

SVL (mm)

Days since 10 May

Fic. 8. Relationship between body size and the date males of R. nigromaculata were first found
calling in 1990.

pip Current Herpetol. 19(1) 2000

No. of nights called

SVL (mm)

Fic. 9. Ralationship between body size and the number of nights of calling in males of R.
nigromaculata in 1990.

へ J
〇

SVL (mm)

Oo
on

60
0 10 20 30 40 50 60

Days since 10 May

Fic. 10. Ralationship between the date of oviposition and the female size of R. nogromaculata in
1990.

SHIMOYAMA—FROG PAIR-FORMATION

nificant tendency for larger females to
spawn earlier than smaller females
(r= —0.566, N=19, p=0.012). This ten-
dency is consistent with the analyses of the
female ovarian cycle of this species (see
Shimoyama, 1993a).

During the course of the study, I found
30 conspecific pairs of R. nigromaculata.
These pairs included individuals without
markings. I observed the sequence of pair-
formation in 13 cases. Of these, 10
(76.9%) were initiated by forced clasping by
males. That is, calling males chased and
clasped the females by force. In the
remaining three cases, a gravid female visit-
ed a calling male and mated with the male.
I could not obtain any evidence of active
mate choice by females. No significant cor-
relation was found between the male and
female sizes in the amplectant pairs
(r=0.046, N=16, p>0.8: Fig. 11).

SVL of males (mm)

60
60 65 70

Jip

Heterospecific pair-formation and hybridiza-
tion

I found a total of 12 heterospecific pairs.
Of these, 11 (91.7 %) were composed of a
male R. nigromaculata and a female R.
porosa brevipoda (Shimoyama, 1999).
Thus, remarkable asymmetry was observed
in the combination of the heterospecific
pairs. I observed the sequence of the heter-
ospecific pair-formation for seven cases. In
all of these heterospecific pairs, male R.
nigromaculata dashed toward, chased, and
clasped gravid females of R. porosa
brevipoda which appeared in the mixed-spe-
cies choruses. Most of the eggs deposited
by these heterospecific pairs hatched nor-
mally (Shimoyama, 1999). So far as I ob-
served in either species, no gravid female
approached calling heterospecific males.

Figure 12 shows the relationship between
the SVL of females and that of males in the
heterospecific pairs. No significant correla-

/ ら 5 80 85

SVL of females (mm)

Fic. 11.
Data from seven seasons were pooled.

Ralationship between the male and female sizes in amplectant pairs of R. nigromaculata.

SVL of males (mm)

50 55 60

Current Herpetol. 19(1) 2000

65 70 15

SVL of females (mm)

Fic. 12. Relationship between the male and female sizes in heterospecific pairs. Dots: pairs com-
posed of a male R. nigromaculata and a female R. porosa brevipoda; cross: a pair composed of a male

R. porosa brevipoda and a female R. nigromaculata.

tion was found between the size of male R.
nigromaculata and that of female R. porosa
brevipoda (r=0.237, N=10, p>0.5).

Speculation on the cause of the asymmetry in
the heterospecific pairing and hybridization

As noted above, gravid females of neither
species seemed to be attracted to the calling
of heterospecific males. This implies that
females of the two species were able to dis-
tinguish heterospecifics from conspecifics by
auditory cues. Nevertheless, a total of 12
cases of heterospecific pairing and spawning
were observed.

In my study site, both sexes of R. porosa
brevipoda were numerically twice as abun-
dant as those of R. nigromaculata. If het-
erospecific pairing occurs at random, the
ratio of the heterospecific pairs with oppo-
site combinations should be 1:1.
However, distinct asymmetry was found in

the combination. Eleven of the 12 heter-
ospecific pairs (91.7%) were composed of
male R. nigromaculata and female R. poro-
sa brevipoda. The ratio of the opposite
combinations of the heterospecific pairs
(11:1) was significantly different from the
expected ratio (1:1; binomial test, p=
0.0063). A similar tendency was also found
under experimental conditions (Shimoya-
ma, 1989). These tendencies suggest that
the asymmetry in the combination of the
heterospecific pairs was not affected by the
difference in the number of individuals of
the two species.

I will examine two ethological factors
which might cause the asymmetry in the
heterospecific pairings and hybridization.

(1) Difference of pairing sequence be-
tween the two species
In R. porosa brevipoda, the majority of

SHIMOYAMA—FROG PAIR-FORMATION

pair-formations was initiated by females.
This indicates that males tend to wait for
the visit of females. On the other hand,
forced clasping by males was the major
pairing pattern in R. nigromaculata (10 out
of the 13 cases). That is, male R.
nigromaculata tended to dash toward and
clasp any moving frog which appeared
nearby. This distinct difference in the pair-
ing sequence seems to be the most im-
portant factor causing the asymmetry in the
heterospecific pairings.

(2) Difference of body size and shape

As shown in Figs. 1 and 7, both male and
female R. nigromaculata were much larger
than R. porosa brevipoda. In addition,
both male and female R. nigromaculata can
move more quickly than R. porosa brevipo-
da, because of the difference in body shape
(see Maeda and Matsui, 1989). It seems
probable that male R. nigromaculata could
easily clasp gravid females of R. porosa
brevipoda. On the other hand, male R.
porosa brevipoda could not easily clasp fe-
male R. nigromaculata, because female R.
nigromaculata are too large and nimble to
be clasped by force.

Influence of hybridization on the two spe-
cies

It is apparent that the costs of the
hybridization for females are more serious
than those for males. Females are forced to
lose whole (or a half) of their gametes in the
season by the hybridization, whereas loss of
male gametes might be less. Hence, R.
porosa brevipoda would suffer more serious
costs than R. nigromaculata by the asym-
metric hybridization. In other words, the
asymmetric hybridization causes one-sided
damage to the population of R. porosa
brevipoda.

Because female hybrids between the two
species are fertile (see Maeda and Matsui,
1989; Matsui, 1996), the female hybrids are
able to produce offspring by means of back-
crosses with males of either species.

Although I have no data on backcrosses,
Nishioka et al. (1981, 1992) showed evi-
dence of introgression between the two spe-
cies inhabiting this basin. The introgres-
sion will also influence the decrease of the
pure genes of R. porosa brevipoda.

ACKNOWLEDGMENTS

I thank Professors Satoshi Yamagishi,
Masafumi Matsui, and Michio Hori, As-
sociate professor Michio Imafuku, Dr.
Akira Mori and the late Professor Hiroaki
Ueda for their helpful advice and sugges-
tions. Thanks are also due to the late
Professor Kenzo Haneda and Professor
Toru Nakamura who led me to the study of
the pond frogs. This study was supported
in part by a Grant-in-Aid (No. 09918012)
from the Ministry of Education, Science
and Culture, Japan.

LITERATURE CITED

ARAK, A. 1983. Male-male competition and
mate choice in anuran amphibians. p.181-210.
In: P. Bateson (ed.), Mate choice. Cambridge
Univ. Press, Cambridge.

INOUE, T. 1979. On the territorial behaviors of
a Japanese pond frog, Rana brevipoda. Jpn.
J. Ecol. 29: 149-161. (in Japanese with English
synopsis)

MAEpA, N. AND M. MarsuI. 1989. Frogs and
Toads of Japan. Bun-ichi Sogo Shuppan,
Tokyo. (in Japanese with English abstract)

MARTIN, P. AND P. BATESON. 1986. Measuring
Behavior. Cambridge Univ. Press, Cam-
bridge.

MArsur, M. 1996. Natural History of the Am-
phibia. University of Tokyo Press, Tokyo. (in
Japanese)

NisHioka, M., M. Sumipa, AND H. OHTANI.
1992. Differentiation of 70 populations in the
Rana nigromaculata group by the methods of
electrophoretic analyses. Sci. Rep. Lab. Am-
phibian Biol. Hiroshima Univ. 11: 1-70.

NisHioka, M., H. UEDA, AND M. Sumipa. 1981.
Genetic variation of five enzymes in Japanese
pond frogs. Sci. Rep. Lab. Amphibian Biol.
Hiroshima Univ. 5: 107-153.

SERIZAWA, T. 1983. Reproductive traits of the

Rana _ nigromaculata-brevipoda complex in
Japan. I. Growth and egg- laying in Tatsuda
and Saya, Aichi Prefecture. Jpn. J. Herpetol.
10: 7-19. (in Japanese with English abstract)
SERIZAWA, T., Y. TANIGAWA, AND S. SERIZAWA.
1990. Reproductive traits of the Rana
nigromaculata-porosa complex in Japan. IV.
Clutch size and ovum size. Jpn. J. Herpetol.
13: 80-86. (in Japanese with English abstract)
SHIMOYAMA, R. 1986a. Distribution and life
history of the Rana nigromaculata species
group in Nagano Prefecture, Japan. Shinano
Kyoiku (1199): 68-73. (in Japanese)
SHIMOYAMA, R. 1986b. Maturity and clutch fre-
quency of female Rana porosa brevipoda in
the northern Ina Basin, Nagano Prefecture,
Japan. Jpn. J. Herpetol. 11: 167-172.
SHIMOYAMA, R. 1989. Why does hybridization
occur between Rana nigromaculata and R.
porosa brevipoda? Jpn. J. Herpetol. 13: 47.

Current Herpetol. 19(1) 2000

(abstract, in Japanese)

SHIMOYAMA, R. 1993a. Female reproductive
traits in a population of the pond frog, Rana
nigromaculata, with prolonged breeding sea-
son. Jpn. J. Herpetol. 15: 37-41.

SHIMOYAMA, R. 1993b. Chorus organization
and male mating behavior in the Japanese
pond frog, Rana porosa brevipoda. J. Ethol.
11: 91-97.

SHIMOYAMA, R. 1996. Sympatric and syn-
chronous breeding by two pond frogs, Rana
porosa brevipoda and Rana nigromaculata.
Jpn. J. Herpetol. 16: 87-93.

SHIMOYAMA, R. 1999. Interspecific interactions
between two Japanese pond frogs, Rana poro-
sa brevipoda and Rana nigromaculata. Jpn. J.
Herpetol. 18: 7-15.

Accepted: 21 March 2000

Current Herpetology 19(1): 27—34., June 2000
© 2000 by The Herpetological Society of Japan

Ant Specialization in Diet of the Narrow-mouthed Toad,

Microhyla ornata, from Amamioshima Island
of the Ryukyu Archipelago

TOSHIAKI HIRAI AND MAsArUMr MATSUI*

Graduate School of Human and Environmental Studies, Kyoto University, Sakyo-ku,

Kyoto 606-8501 JAPAN

Abstract: Microhyla ornata consumed numerous ants, representing 77.1% in
number and 44.6% in volume of the diet. The toad took ants in higher
proportion than were present in the surrounding environment, and therefore,
could be viewed as an ant specialized predator. Ants were the most
numerously consumed prey in both spring and summer, while beetles and
woodlice were less frequently taken in summer. Females have a larger body
and wider mouth than males, and consumed significantly larger prey in maxi-
mum size than did males. However, mean prey size, and frequencies of oc-
currence for all prey taxa did not differ significantly between the sexes. These
results suggest that the sexes do not differ in their use of food resources despite

their morphological differences.

Key words:
chipelago; Gape-limited predator

INTRODUCTION

The Anuran assemblage in the Ryukyu
Archipelago (excepting Osumi Islands) is
unique and more highly diversified than that
found in the adjacent mainland of Japan.
While 21 anuran species/subspecies occur in
mainland Japan, 20 other species are dis-
tributed in the Ryukyu Archipelago, and 14
of them are endemic to the archipelago
(Maeda and Matsui, 1999).

Quite a few comprehensive taxonomic
and biogeographical studies have been
made on anurans in the Ryukyu Archipela-

* Corresponding author. Tel: +81-75-753-
6846; Fax: +81-75-753-2891.

E-mail address: fumi@zoo.zool.kyoto-u.ac.jp
(M. Matsui)

Microhyla ornata; Ant-specialists; Prey availability; Ryukyu Ar-

go (e.g., Matsui, 1994; Ota, 1998), but eco-
logical studies are much more limited, and
mostly confined to reproduction (e.g., Ut-
sunomiya, 1980, 1989). A preliminary
report by Okochi and Katsuren (1989) is the
only information available about diet of
anurans on Okinawajima Island.

Microhyla ornata ranges throughout the
Ryukyu Archipelago from Amamioshima
southward through China to Southeast Asia
and India (Frost, 1985), and inhabits vari-
ous habitats from lowlands to montane
regions (Maeda and Matsui, 1999). The
toad is presumed to be specialized for eating
ants or termites because of its small body
and narrow mouth (Maeda and Matsui,
1999). However, detailed quantitative stu-
dies on the food habits of this species have
not been done so far.

Berry (1965) reported that ants predomi-
nated in the diet of two congeneric species,
M. butleri and M. heymonsii from Singa-
pore. Many dendrobatid and bufonid spe-
cies are also well-known for _ eating
numerous ants, and are called ant specialists
because they take ants in higher proportion
than are found in the environment (Toft,
1980, 1981).

Toft (1985), in reviewing resource par-
titioning studies in amphibians and reptiles,
considered that food partitioning plays an
important role in the organization of adult
anuran communities. Therefore,
knowledge of food habits of each com-
munity member might provide pivotal in-
sights into the factors that are responsible
for frog community structures in the Ryu-
kyu Archipelago.

In order to obtain information on prey
selection by the narrow-mouthed toad, M.
ornata, we conducted field work on
Amamioshima which is near the northern
limits of its range.

MATERIALS AND METHODS

For the diet study, we collected toads in
evergreen forests of the outskirts of Naze
city on Amamioshima Is. (28°22’N,
129?31 E). We made collections at night
(2100-2300 h) on 6 April (spring) and 12
July (summer) of 1998. We captured all in-
dividuals encountered on the forest floor,
and immediately fixed them in 10% buffered
formalin to preserve stomach contents with
minimum digestion.

In order to estimate prey availability in
the habitat of the toads, we sampled leaf
litter invertebrates by two _ different
methods. In spring, we set pit-fall traps
near the temporal pool where toads were
abundant. Fifteen plastic cups (90mm in
diameter and 130 mm in depth) were set in
the ground at about 2m intervals for two
nights from 7 to 9 April. A small quantity
of ethylene glycol was put into each cup to
preserve samples. In summer, at the same

Current Herpetol. 19(1) 2000

time as the collection of toads, we collected
four 0.5x0.5m quadrate samples of leaf
litter and brought them to the laboratory.
After large or inactive animals were re-
moved from these samples, the remaining
ones were extracted by Tullugren’s funnel
method (Aoki, 1973). These potential prey
invertebrates were stored in ethylene glycol
for later analyses.

Stomach contents were removed by dis-
section of toads, and preserved in 10%
buffered formalin for later analyses. For
each toad, snout-vent length (SVL) and
mouth width (MW) were measured with a
caliper to the nearest 0.1 mm.

We identified stomach contents and
potential prey to the level of class or order
except for Hymenoptera, which was clas-
sified into Formicidae and non-Formicidae.
For holometabolous insects, larvae and
adults were separately treated. The occur-
rence of plant materials or minerals was
recorded for each stomach. Details about
measurement of stomach contents are given
in Hirai and Matsui (1999).

To detect seasonal variation in the diet,
we compared the presence or absence of
each prey taxon between spring and summer
by Fisher’s exact probability test. Next, we
examined the relationships between prey
availability and diet composition by cal-
culating Kendall’s rank _ correlation
coefficients (t). In this analysis, we used
only taxa that were commonly found in
both potential prey samples and _ the
stomach contents because prey taxa found
only in one of these might have large sam-
pling errors. We presumed that the exclu-
sion of these uncommon prey taxa would
not affect the result of analyses because the
common prey taxa accounted for more than
75% of both diet and prey samples.
Moreover, we tested the sexual difference in
the diet by comparing the presence or ab-
sence of each prey taxon by Fisher’s exact
probability test. In additition, we quan-
tified dietary overlap between sexes by cal-
culating simple similarity indices (Schoener,

HIRAI & MATSUI—ANT SPECIALIZATION BY A MICROHYLID 29

1968):

Oat 0S) Pig P 3, |
based on proportion of prey taxa (i) in diet
of two different sexes (x and y). We calcu-
lated both numeric and volumetric overlap
in this analysis.

RESULTS

Diet composition

We identified 1234 prey items extracted
from 55 stomachs (41 males, 14 females) of
56 individuals captured; the remaining
stomach from an immature female was
empty. Prey items included five arthropod
classes (Arachnida, Crustacea, Diplopoda,
Chilopoda, and Insecta), of which Insecta
contained eight orders (Table 1). Ants
(Formicidae) were consumed by all individ-
ual toads with stomach contents, and
predominated in the diet, representing
77.1% by number, and 44.6% by volume.
The next most frequently consumed prey
taxon was beetles (Coleoptera; 67.3%), but
they made up only 6.4% by number, and
16.9% by volume. The other prey taxa
constituted minute fractions, making up
less than 5% by number, and 15% by
volume. Plant (vegetable scraps) and
mineral materials (pebbles and dirt) oc-
curred in 47.3% and 1.8%, respectively, of
the stomachs.

Seasonal variation in diet

The toads appeared to take more prey in
spring (Mean + SE= 26.1 +3.9) than in sum-
mer (16.5+2.7), but the difference was not
significant (U-test, p >0.05). However, the
volume of stomach contents differed
seasonally, and almost three times more
food was ingested in spring (59.2+8.1 mm?)
than in summer (20.8 土 4.0 mm?)) (U-test,
p<0.01).

Ants constituted the bulk of the diet nu-
merically and volumetrically, and their
proportions varied little seasonally (Table
2). Among 10 prey taxa commonly con-
sumed in both seasons, all prey taxa except

TABLE 1. Diet composition (in %) of Micro-
hyla ornata (1234 prey from 55 individuals, total
volume 2448.3 mm3 う ). Abbreviations: F=fre-
quency of occurrence; N=numeric proportion;
V=volumetric proportion

Prey taxa F N V
Insecta
Hymenoptera
Formicidae 100.0 IAFL 44.6

non-Formicidae Sy) 0.2 0:5

Coleoptera 67.3 6.4 16.9
larvae 3:6 0.3 4.1
Diptera 14.6 10 0.8
larvae 10.9 1.0 4.4
Lepidoptera 1.8 Or 02
larvae eo <0nl 0.9
Hemiptera 20.0 i 3:2
Isoptera 3.6 4.7 1.4
Orthoptera 1.S 6«< 051 0.3
Collembola 21.8 j PS) 0.2
Arachnida
Araneae 10.9 0.7 at
Pseudoscorpiones ON 0
Acarina 218 25 0.5
Crustacea
Isopoda 20.0 eS MISS3
Chilopoda i 2e9/ 0.7 2.6
Diplopoda 21.8 ie? a |
Plant materials 47.3 ーー ーー
Minerals 1.8 ーー 一

for ants and wasps (non-formicids) were
found more frequently in spring than in
summer, but only two prey taxa, beetles and
woodlice (Isopoda), differed significantly in
fequency of occurrence (Fisher’s exact
probability test, p<0.01 for Coleoptera;
p<0.05 for Isopoda). Volumetric contri-
bution by these two prey taxa was great in
spring, but it showed striking decrease in
summer. By contrast, termites (Isoptera)
made up a larger numerical proportion in
summer because of two individuals (9.5%
in frequency) that contained unusually
many termites (Table 2). Frequency of oc-

Current Herpetol. 19(1) 2000

TABLE 2. Dietary comparison of Microhyla ornata between spring (887 prey from 34 individuals,
total volume 2012.4 mm?) and summer (347 prey from 21 individuals, total volume 435.9 mm3). See

Table 1 for abbreviations.

F N V
Prey taxa Spring Summer Spring Summer Spring Summer
Insecta
Hymenoptera
Formicidae 100.0 100.0 79.8 70.0 45.0 43.1
non-Formicidae 2.9 OFS 0.1 0.6 0.3 1.6
Coleoptera 82.4 42.9 7 が 9/ 3.2 18.4 9.5
larvae 0 9.5 0 1 0 2.3 22
Diptera IM 9.5 | 0.6 0.7 1.0
larvae ea 0 1.4 0 5.4 0
Lepidoptera 0 4.8 0 0.6 0 1.0
larvae 2.9 0 0.1 0 1.1 0
Hemiptera 23.5 14.3 0.9 1.4 3.4 DES
Isoptera 0 9:5 0 16.7 0 7
Orthoptera 0 4.8 0 0.3 0 1.7
Collembola 26.5 14.3 jee 0.9 0.2 0.1
Arachnida
Araneae 14.7 4.8 0.8 0.3 1.3 ぐ 0.1
Pseudoscorpiones 0 4.8 0 0.3 0 <0.1
Acarina 26.5 14.3 2.6 の 88 0.3 1.4
Crustacea
Isopoda 29.4 4.8 1.6 0.6 12.8 4.0
Chilopoda 20.6 0 1.0 0 322 0
Diplopoda 26.5 14.3 1.2 i 7.8 3.4
Plant materials 26.5 90.5 ーー — — —
Minerals 2.9 0 — — ーー ーー

currence of plant materials markedly in-
creased from spring (26.5%) to summer
(90.5%), and differed significantly between
the seasons (Fisher’s exact probability test,
p<0.01).

Prey selection

A total of 20 potential prey invertebrates
were collected when samples in spring and
summer were combined (Table 3). Among
these, larval caddisflies (Trichoptera), ear-
wigs (Dermaptera), gastropod snails, and
earthworms (Oligochaeta) were not found
in the toad stomachs. Conversely, moths
(Lepidoptera) and their larvae were not

sampled from the surrounding habitat.
Specifically in spring, 11 of 14 potential
prey invertebrates sampled from the en-
vironment were found in diet composition.
Ants seemed to be the most readily available
prey for toads, and were consumed in much
higher proportion than those found in the
environment. However, the next most
abundant possible prey, collembolans, were
consumed much less in proportion by toads.
Consequently, there was no significant cor-
relation between prey availability in the
habitat and diet composition (t=0.278,
p>0.05). In summer, 12 of 17 potential
prey invertebrate taxa were actually found

HIRAI & MATSUI—ANT SPECIALIZATION BY A MICROHYLID

TABLE 3.

Comparison of diet composition of M. ornata with prey availability in the environment

assessed by pitfall traps in spring and funnel traps in summer.

Spring Summer
Diet Environment Diet Environment
Prey taxa n % n % n % n %

Formicidae 708 79.8 97 41.6 243 70.0 40 10.6
non-Formicidae 1 0.1 1 0.4 2 0.6 0 0
Coleoptera 68 ed. 10 4.3 11 3 4 il

larvae 0 0 0 0 4 eZ 15 4.0
Diptera 10 iol 26 eZ 2/ 0.6 4 |

larvae 12 1.4 1 0.4 0 0 1 0.3
Lepidoptera 0 0 0 0 2 0.6 0 0

larvae 1 0.1 0 0 0 0 0 0
Trichoptera larvae 0 0 0 0 0 0 1 0.3
Hemiptera 8 0.9 2 0.9 5 1.4 7) 0.5
Isoptera 0 0 6 2.6 58 16.7 8 Zh
Dermaptera 0 0 0 0 0 0 1 0.3
Orthoptera 0 0 0 0 1 0.3 1 0.3
Collembola 15 Nea 35 15.0 3 0.9 1 0.3
Araneae 7 0.8 17 TS 1 0.3 ウ 0.5
Pseudoscorpiones 0 0 1 0.4 1 0.3 0 0
Acarina 23 2.6 3 1.3 8 P18) 161 42.8
Isopoda 14 1.6 11 4.7 ウ 0.6 35 93
Chilopoda 9 1.0 0 0 0 0 1 0.3
Diplopoda 11 je 17 es 4 12 11 2.9
Gastropoda 0 0 6 2.6 0 0 0 0
Oligocheata 0 0 0 0 0 0 88 23.4

in stomachs of toads. As was found in spr-
ing, the proportion of ants in the diet was
larger than that found in the environment.
Instead, mites, the most abundant prey in
the environment, were consumed by few
toads. We could not detect a significant
correlation between prey availability and
diet composition (t=0.381, p >0.05).

Comparisons between sexes

Females (mean + SE=30.5 +0.6 mm, ran-
ge=25.6—33.0 mm) were significantly larg-
er in SVL than males (26.3+0.2 mm, 22.1-
29.6mm) (U-test, p<0.01). Females also
had a significantly wider mouth (8.3+
0.1mm, 7.5-9.3mm) than males (7.34
0.1mm, 6.1-8.5mm) (p<0.01). Sexual

difference was highly significant in maxi-
mum prey size (females: 19.9+4.1 mm;
males: 12.0 土 2.2 mm) (p<0.01), but was
not significant in mean (females: 3.04
0.5mm, males: 1.90.2 mm) or minimum
prey size (females: 0.6+0.2mm, males:
0.3+0.04mm) (p>0.05 for both) due to
their large variations. Similarly, neither
the number (females: 28.7+8.0, males:
20.3 土 2.4) nor the volume (females:
66.8+16.8 mm?, males: 36.9+4.8 mm?) of
stomach contents differed significantly be-
tween the sexes (p > 0.05 for both).

Diet compositions did not differ markedly
between females and males as suggested
by high dietary overlap (similarity in-
dices=0.85 in number and 0.70 in volume:

Current Herpetol. 19(1) 2000

TABLE 4. Dietary comparison of female (402 prey from 14 individuals, total volume 934.6 mm)
and male (832 prey from 41 individuals, total volume 1513.7 mm?) Microhyla ornata. See Table 1 for

abbreviations.
F N V
Prey taxa Female Male Female Male Female Male
Insecta
Hymenoptera
Formicidae 100.0 100.0 86.1 WZ S726 36.5
non-Formicidae ial 4.9 0.3 0.2 0.7 0.5
Coleoptera 575l TOT. 3.0 8.1 9.3 2S
larvae el 2.4 0.3 0.4 2.6 5.1
Diptera Tal WA 0.3 1.3 0.9 0.7
larvae 21.4 ES: クウ の 0.4 8.0 クン
Lepidoptera 0 2.4 0 0.2 0 0.3
larvae Al 0 0.3 0 2.4 0
Hemiptera 14.3 22.0 0.8 h.2 1.5 4.3
Isoptera 0 4.9 0 7.0 0 Ze
Orthoptera 0 2.4 0 0.1 0 0.5
Collembola Tal 26.8 0.5 1.9 <0.1 0.2
Arachnida
Araneae 21.4 ee) 1.0 0.5 ts] 0.8
Pseudoscorpiones qa 0 2.0 0 ぐ 0.1 0
Acarina 42.9 14.6 0.3 2.8 0.3 0.6
Crustacea
Isopoda 14.3 22.0 1.0 1.4 8.4 13.1
Chilopoda 14.3 12ND, 0.8 0.7 1.2 3.5
Diplopoda 28.6 19:5 shes) 1.1 の 内 9 8.2
Plant materials 42.9 S320 — — — —
Minerals od 0 — — — —
Table 4). Indeed, frequency of occurrence tion than those available from the environ-

of all prey taxa did not differ significantly
between the sexes (Fisher’s exact probability
test, p>0O.05 for all pray taxa). Ants
predominated in their diet, both numerical-
ly and volumetrically (Table 4).

DISCUSSION

Ants were consumed by all individuals of
M. ornata studied, and comprised more
than 70% of the total prey items. Follow-
ing Simon and Toft’s (1991) definition, M.
ornata is regarded as an ant specialist be-
cause they consumed ants higher in propor-

ment.

Berry (1965) reported that ants occurred
in more than 94.9% of stomachs, and made
up 86-87% of the total prey items in the diet
of two microhylids, M. butleri and M. hey-
monsii from Singapore. Results of our
study in M. ornata, i.e., ants occurring in
all stomachs, and making up 77.1% of total
prey items, are consistent with Berry’s
(1965) observation. Therefore, the food
habits of this genus would be characterized
by eating numerous ants.

Seasonal variation in diet was conspicu-
ous in beetles and woodlice. In spring,

HIRAI & MATSUI—ANT SPECIALIZATION BY A MICROHYLID 33

these prey taxa were consumed more fre-
quently, and made up relatively large
proportions by volume. Fewer chances of
consuming these prey might have caused a
decrease in the amount of food ingested
during summer. Instead, plant materials
were found more frequently in summer. A
ranid frog, Rana hexadactyla, has been
reported to supplement energy gain by con-
suming plant materials (Das, 1996), and M.
ornata might also consume plant materials
frequently in summer so as to compensate
for reduction of animal food consumption.

Sexual dimorphism in body size and
mouth width was observed in Microhyla or-
nata, and both the variables were larger in
females than in males. Generally, anurans
are gape-limited in predation, and size of
consumable prey is regulated by body size
or mouth width (e.g., Kramek, 1972; Toft,
1980). Hence, such sexual dimorphism in
morphology may help to alleviate potential
competitive interactions by partitioning
food resources. In fact, females of Rana
cancrivora with larger body and wider
mouth than males were reported to con-
sume larger prey than males (Premo and
Atmowidjojo, 1987). In our observations,
where maximum prey size was larger in fe-
males suggests that consumable prey size in
M. ornata is also determined by the mouth
width. However, the sexes did not differ in
mean or minimum prey size. Further, they
were similar in frequency of occurrence for
all prey taxa. These lines of evidence indi-
cate that food resources were not par-
titioned between the sexes. High dietary
overlap of females and males also seems to
support this assumption.

Among anurans occurring in the Ryukyu
Archipelago, diet data for four ranid spe-
cies, Rana namiyei, R. narina, R. ishika-
wae, and R. holsti from Okinawajima Is-
land, are available (Okochi and Katsuren,
1989). These four species are relatively
large, reaching 55 to 120mm in SVL, and
consume few ants. Therefore, ant speciali-
zation in M. ornata of less than 30 mm in

SVL might be partially responsible for the
coexistence with larger anuran species.
Anuran fauna in the subtropical Ryukyu
Archipelago is characterized by the presence
of many endemic species, and by greater
species diversity than in temperate mainland
Japan (Matsui, 1996).

In the tropics where species diversity is
greater than in temperate and subtropical
regions, and anurans have specialized their
various ecological attributes (Duellman and
Trueb, 1986), diet specialization has been
demonstrated to be important in structuring
an anuran community (Toft, 1980). There-
fore, ant specialization in M. ornata might
be playing a great role in structuring anuran
communities in the Ryukyu Archipelago as
well. Further studies based on community
ecology are necessary to evaluate the impor-
tance of the role of food specialization.

LITERATURE CITED

AOKI, J. 1973. Soil Zoology. Hokuryukan,
Tokyo. (in Japanese)

BERRY, P. Y. 1965. The diet of some Singapore
Anura (Amphibia). Proc. Zool. Soc. London
144: 163-174.

Das, I. 1996. Folivory and seasonal changes in
diet in Rana hexadactyla (Anura: Ranidae). J.
Zool. London 238: 785-794.

DUELLMAN, W. E. AND L. TRUEB. 1986. Biology
of Amphibians. McGraw-Hill Book Co., New
York.

Frost, D. R. (ed.). 1985. Amphibian Species of
the World: A Taxonomic and Geographical
Reference. Allen Press, Lawrence, Kansas.
l=Vel- (O2D-

Hiral, T. AND M. MArsur. 1999. Feeding habits
of the pond frog, Rana nigromaculata, in-
habiting rice fields in Kyoto, Japan. Copeia
1999(4): 940-947.

KRAMEK, W.C. 1972. Food of the frog Rana
septentrionalis in New York. Copeia 1972:
390-392.

Maepba, N. AND M. MArsUr. 1999. Frogs and
Toads of Japan. Rev. Ed. Bun-ichi Sogo
Shuppan, Tokyo. (in Japanese with English
abstract)

Matsul1, M. 1994. A taxonomic study of the
Rana narina complex, with description of

Current Herpetol. 19(1) 2000

three new species (Amphibia: Ranidae). Zool.
J. Linn. Soc. 111: 385-415.

MArsur, M. 1996. Natural History of the Am-
phibia. University of Tokyo Press, Tokyo. (in
Japanese)

Oxoculi, I. AND S. KATSUREN. 1989. Food
habits in four species of Okinawan frogs.
p. 405-412. In: M. Matsui, T. Hikida, and
R.C. Goris (eds.), Current Herpetology in
East Asia. Herpetol. Soc. Jpn., Kyoto.

Ota, H. 1998. Geographic patterns of ende-
mism and speciation in amphibians and rep-
tiles of the Ryukyu Archipelago, Japan, with
special reference to their paleogeographical
implications. Res. Popul. Ecol. 40: 189-204.

PREMO, D. B. AND A. H. ATMOWIDJOJO. 1987.
Dietary patterns of the crab eating frog Rana
cancrivora in west Java. Herpetologica 43: 1-
6.

SCHOENER, T. W. 1968. The Anolis lizards of
Bimini: resource partitioning in a complex
fauna. Ecology 49: 704-726.

SIMON。 M.P. ANDp C.A. Tort. 1991. Diet
specialization in small vertebrates: mite-eating
in frogs. Oikos 61: 263-278.

Tort, C. A. 1980. Feeding ecology of thirteen
syntopic species of anurans in a seasonal trop-
ical environment. Oecologia 45: 131-141.

TorT, C. A. 1981. Feeding ecology of Panama-
nian litter anurans: patterns in diet and forag-
ing mode. J. Herpetol. 15: 139-144.

Tort, C. A. 1985. Resource partitioning in am-
phibians and reptiles. Copeia 1985: 1-21.

UtsunomiyA, T. 1980. Reproduction of Rana
ishikawae on Amamioshima Island. Nippon
Herpetol. J. 17: 17-18. (in Japanese)

UrsUNowryA, T. 1989. Five endemic frog spe-
cies of the Ryukyu Archipelago. p. 199-204.
In: M. Matsui, T. Hikida, and R.C. Goris
(eds.), Current Herpetology in East Asia. Her-
petol. Soc. Jpn., Kyoto.

Accepted: 10 April 2000

Current Herpetology 19(1): 35—40., June 2000
© 2000 by The Herpetological Society of Japan

Nomenclatural History and Rediscovery of
Rhacophorus lateralis Boulenger, 1883
(Amphibia: Rhacophoridae)

INDRANEIL DAS

Institute of Biodiversity and Environmental Conservation,
Universiti Malaysia Sarawak, 94300, Kota Samarahan,
Sarawak, MALAYSIA

Abstract: The poorly known south-west Indian rhacophorid, Rhacophorus
lateralis Boulenger, 1883, known from a unique holotype in the BMNH, is
redescribed based on two adult females, from South Coorg, Karnataka State,
south-western India. The species is listed in recent lists as valid, despite an
attempted synonymy with Rhacophorus malabaricus Jerdon, 1870, by Wolf
(1936). The species is diagnosed by the following suite of characters: skin of
forehead free; dorsum dark brown with a pair of yellow lines that run from the
region around the nostrils, over the eyelids, along the sides of the body, ter-
minating in the inguinal region; small adult body size (SVL to 32.8mm). The
species is illustrated for the first time.

Key words:
Redescription; Western Ghats; India

INTRODUCTION

Rhacophorus lateralis was described by
Boulenger (1883) based on a unique holo-
type (BMNH 82.2.10.75) from “Malabar”
(at present in southern Kerala State, south-
western India) that was collected by Colonel
Richard Henry Beddome. No further
specimens of this species have come to light
since the original description, despite
numerous surveys of the hill ranges of
southern India that are referred to as the
Western Ghats (see Mani, 1974, for a
description and Dutta, 1997, for a bib-

Tel: +60-82-671000 (ext. 247); Fax: 十 60-
82-671903.
E-mail address: idas@mailhost.unimas.my

Rhacophorus lateralis; Nomenclatural history; Rediscovery;

liography of published works on amphibi-
ans of the region). Several workers listed
the species as valid: Boulenger (1890: 473);
Ahl (1931: 165, as Rhacophorus [Rhaco-
phorus] lateralis); Inger and Dutta (1986),
Daniel and Sekar (1989), Daniels (1992),
Dutta (1997: 102), and Das and Dutta
(1998). On the other hand, the species is
not listed by Gorham (1974), Frost (1985),
Dubois (1986), or Duellman (1993), perhaps
following Wolf (1936: 214) who considered
it as possibly synonymous with Rhacopho-
rus reinwaratii malabaricus (= R. mala-
baricus Jerdon, 1870). The taxon has never
been illustrated.

The collection of two adult females and
an unsexed metamorph assignable to
Rhacophorus lateralis Boulenger, 1883,

Current Herpetol. 19(1) 2000

from Lakunda Estate (12?03 18"N:
76°02’ 62 E), Virajpet District, Srimangala
Nadu, South Coorg (Kodagu) District, Kar-
nataka State, south-western India, provides
the opportunity to redescribe the species on
the basis of the two adult females, illustrate
one of these, and announce the rediscovery
of the species after over a century.

METHODS

Measurements were taken with a
Mitutoyo™ dial caliper (to the nearest
0.1mm) from specimens preserved in
ethanol. The following measurements were
taken: snout-vent length, SVL (from tip of
snout to vent); tibia length, TBL (distance
between surface of knee to surface of heel,
with both tibia and tarsus flexed); axilla to

Fic. 1. Adult female Rhacophorus lateralis (ZSI A9071).

groin distance, A-G (distance between
posterior edge of forelimb at its insertion to
body to anterior edge of hind limb at its in-
sertion to body); head length, HL (distance
between angle of jaws and snout-tip); head
width, HW (measured at angle of jaws);
head depth, HD (greatest transverse depth
of the head, taken posterior to orbital
region); eye diameter, ED (diameter of the
orbit); eye to tympanum distance, E-T (dis-
tance between posterior-most point of orbit
and anterior-most edge of tympanum); up-
per eyelid width, UE (greatest width of up-
per eyelid); interorbital width, IO (least dis-
tance between upper eyelids); internarial
distance, IN (distance between nostrils); eye
to snout-tip distance, E-S (distance between
anterior-most point of orbit to tip of snout);
eye to nostril distance, E-N (distance be-

Marker = 15Q12mm.

DAS—REDISCOVERY OF RHACOPHORUS LATERALIS ます /

tween anterior-most point of orbit and nos-
tril); horizontal tympanum _ diameter,
HTYD (diameter of left tympanum, taken
across hozontal plane); vertical tympanum
diameter, VT YD (diameter of left tympa-
num, taken across vertical plane); and di-
ameter of disk on Finger III, FIIID (greatest
width of terminal disk on Finger III).
Lengths of digits were measured from the
base of the web to the distal tips of the
digits. Institutional abbreviations follow
Leviton et al. (1985).

REDESCRIPTION OF RHACOPHORIS
LATERALIS BOULENGER, 1883

Redescription (based on ZSI A9071-72,
both adult females)

SVL 29.5 and 32.8 mm (subsequent ratios
in the same sequence); habitus slender (Fig.
1); head relatively short (HL/SVL ratio 0.27
and 0.26) and broad (HW/SVL ratio 0.31
and 0.30), its width greater than its length
(HW/HL ratio 1.11 and 1.16); snout short,
obtusely pointed, projecting a little beyond
level of mandibles; nostril closer to tip of

TABLE 1.
south-western India (see text for details).

snout than to eye (E-N/E-S ratio 0.58 and
0.57). Canthus rostralis flattened in trans-
verse section; lores slightly concave. Eyes
large (ED/HL ratio 0.63 and 0.49); eye di-
ameter greater than eye-nostril distance
(ED/E-N ratio 2.22 and 1.64); interorbital
distance greater than width of upper eyelid
(1O/UE ratio 1.59 and 2.64). Supratym-
panic fold absent. Tympanum distinct, its
diameter greatest vertically (HTYD/VTYD
ratio 0.94 and 0.80), less than eye diameter
(HTYD/ED ratio 0.29 and 0.39), situated
posteroventrally to level of eyes, at a little
distance from the posterior corner of the
eyes. Nostrils dorsolaterally oriented,
oval. Vomerine teeth in two short, oblique
series between choanae. Inner margin of
mandible with a weak w-shaped notch an-
teriorly. Choanae oval, separated from
each other by a distance over five times
greater their width. Tongue large, smooth,
lacking median conical lingual papilla,
elongate, bifid and free posteriorly.
Forelimbs relatively short; tips of fingers
dilated into large, flattened, rounded disks
(Fig. 2a) with circummarginal grooves; the

Morphometric data (in mm) on two adult female Rhacophorus lateralis from Karnataka,

Snout-vent length
Axilla-groin distance

Head length

Head width

Head depth

Eye diameter

Upper eyelid width
Interorbital distance
Internarial distance
Eye-snout-tip distance
Eye-nostril distance
Eye-tympanum distance
Tibia length

Disk diameter finger III
Horizontal tympanum diameter
Vertical tympanum diameter

ZSI A9071 ZSI A9072
29.5 32.0
14.1 18.0

8.1 6S9
9.0 9.6
5.6 323
5.1 4.1
Dee ジジ
9 5.8
2.8 2S
4.0 4.4
2g) 2.5
0.6 0.7
16.3 17.8
ed) [25
1.5 1.6
1.6 2.0

Current Herpetol. 19(1) 2000

Fic. 2. Fore and hind limb of Rhacophorus lateralis (ZSI A9071), showing subarticular tubercles
on venter of hand (2a), and on ventral surface of foot (2b). Markers = 5mm.

largest disk (on Finger III) approximately
equal to horizontal diameter of tympanum
(FIIID/HTYD ratio 1.00 and 0.94). Fin-
gers broadly webbed, most extensive web-
bing between Fingers I and II not reaching
proximal subarticular tubercles. All fingers
with dermal fringes on inner and outer
aspects. No dermal fringe on elbow.
Relative lengths of fingers:3 > 4 > 2 > 1.

Hind limbs relatively long; tibia long
(TBL/SVL ratio 0.55 and 0.56); tips of toes
dilated into flattened disks (Fig. 2b) with

TABLE 2.

circummarginal groove, are slightly smaller
than those on fingers. Webbing on Toe I
between distal subarticular tubercle and
digit tip; on Toe II to base of disks on outer
edge and to slightly beyond distal subarticu-
lar tubercle on inner edge; on Toe III to
base of disks on outer edge and to beyond
distal subarticular tubercle on inner edge;
on Toe IV, to base of disk on outer edge
and to the distal subarticular tubercle, as a
broad web, reaching the tip as a narrow
sheath in the inner; and on Toe V, to base

Lengths of digits (in mm) of left hand and foot of two adult female Rhacophorus later-

alis from Karnataka, south-western India (see text for details).

Fingers
1 2 3
ZSI A9071 3.4 4.4 6.4
ZSI A9072 4.2 4.9 6.8

6.0
6.2

Toes
1 Zz 3 4 5
3.4 5.0 8.0 10.8 8
9 5.6 8.5 12.3 9.7

DAS—REDISCOVERY OF RHACOPHORUS LATERALIS 39

of disks. Toes I and V have dermal fringes
on outer edges. Distinct, elongated inner
metatarsal tubercle, outer metatarsal tuber-
cle absent. Relative lengths of toes: 4 > 5
Pa 2-1.

Dorsum smooth, lacking tubercles. Out-
er edge of upper eyelids faintly granular.
Surface of throat, pectoral and abdominal
regions smooth. Undersurface of fore-
limbs and the undersurface of thighs
smooth; trailing edge of hind limbs without
rounded tubercles. Cloacal opening direct-
ed posteroventrally, close to lower level of
thighs.

Colouration (in preservative)

ZSI A9071 has a dark chestnut dorsum,
with a pale yellow stripe that commences
from above the nostrils, running over the
upper eyelids, to over the tympanum and
along the sides of the body, terminating in
the inguinal region. Upper surfaces of
thigh and shank with narrow dark brown
stripe, bounded by more extensive areas of
pale yellow. Dorsum patterned with fine
dark dots. Forearm and thigh with narrow
dark brown bars on the dorsum. Upper
lip, upper arm, digits, undersurface of
limbs and body unpatterned pale yellow.
The posterior surface of thigh unpatterned
yellow. The heel with a pale yellow, sinous
marking. In life, the field notes of the col-
lectors indicate that the frogs were green
dorsally, dotted with blue. ZSI A9072-73
are discoloured, although both show the
pale lateral stripes and the finely dark-band-
ed dorsal surface of limbs.

Size variation and other notes

The SVL of the holotype was reported to
be 31 mm in the original description, while
the two adult females being reported here
are 29.5 and 32.8 mm; the smallest individ-
ual (ZSI A9073, a metamorph with a pale
lateral stripe and the vestige of a tail) is
16.7mm. Other measurements are in Ta-
bles 1-2. No males are represented in the
present series, and the small size of the adult

Fic. 3. Radiograph of adult female Rhaco-
phorus lateralis (ZSI A9071).

females (with developed ovaries) is remark-
able among members of the genus. Skin
over the cranium is free, not coossified to
frontoparietal, nasal, or squamosal bones.
Figure 3 shows radiographs of ZSI A9071.

Comparisons

Because the species has been considered
synonymous with Rhacophorus malabari-
cus by some workers, R. /ateralis is here
compared with congenerics from _ the
Western Ghats. Only opposing suites of
characters are listed. R. malabaricus Jer-
don, 1870: dorsum green, lacking light
lateral stripes; digits webbed to disks; and
SVL to 48.3 mm; R. calcadensis Ahl, 1927,
dorsum reddish-brown, lacking light lateral
stripes; digits webbed to disks; tympanum
40% eye diameter; and SVL to 48.3 mm; R.
pseudomalabaricus Vasudevan and Dutta,
2000, dorsum green, lacking light lateral
stripes; tympanum indistinct; head length
equals width; and SVL 66.8mm; and R.
lateralis Boulenger, 1883; dorsum brown,
lacking distinct light lateral stripes; and
tympanum half eye diameter.

Current Herpetol. 19(1) 2000

COLLECTION NOTES

The adult females were collected on 19
and 29 July 1998. The metamorph was dis-
covered on a leaf above a tank on 3 August
1998. The locality is a moist evergreen
forest within Lakunda Estate, Virajpet Dis-
trict, Srimangala Nadu, South Coorg
(Kodagu) District, Karnataka State, south-
western India. The discovery of the meta-
morph in the month of August indicates
that the species breeds during the summer
(Southwest) Monsoons. Rhacophorus
malabaricus was taken alongside R. lateralis
at this locality.

ACKNOWLEDGMENTS

I thank Daniel Bennett and members of
the Aberdeen University Expedition to the
Western Ghats, July-August, 1998, for col-
lecting the specimens of Rhacophorus later-
alis referred to, and making these and as-
sociated data available to me. J. R. B. Al-
fred and S. K. Chanda provided permission
and facilities at the ZSI. Radiograph of
ZSI A9071 was prepared by Ng Heok Hee,
Raffles Museum of Biodiversity Research,
National University of Singapore, where I
thank Peter Ng, C.M. Yang, and Kelvin
K. P. Lim for facilities. A visit to the NUS
was supported by a grant (UNIMAS 120/98
[9]) from Universiti Malaysia Sarawak. Fi-
nally, grateful thanks are due to an anony-
mous reviewer for comments on a draft
manuscript.

LITERATURE CITED

AHL, E. 1931. Anura III. Polypedatidae. Das
Tierreich 55: i-xvi + 1-477.

BOULENGER, G.A. 1883. Description of new
species of reptiles and batrachians in the Brit-
ish Museum. Ann. Mag. Nat. Hist. (5) 12:
161-167.

BOULENGER, G. A. 1890. The Fauna of British
India, Including Ceylon and Burma. Reptilia
and Batrachia. Taylor and Francis, London.
XVlil + 541 p.

DANIEL, J.C. AND A.G. SEKAR. 1989. Field
guide to the amphibians of Western India.
Part IV. J. Bombay Nat. Hist. Soc. 86: 194-
202.

DANIELS, R. J. R. 1992. Geographical distribu-
tion patterns of amphibians in the Western
Ghats, India. J. Biogeogr. 19: 521-529.

Das, 1. ANDS. K. DuTTA. 1998. Checklist of the
amphibians of India, with English common
names. Hamadryad 23(1): 63-68.

Dusols, A. 1986. Miscellanea taxinomica
batrachologia (I). Alytes 5(1/2): 7-95.

DUELLMAN, W.E. 1993. Amphibian species of
the world: Additions and corrections. Univ.
Kansas Mus. Nat. Hist. Sp. Publ. (21): i-iii +
1-372.

Dutta, S.K. 1997. Amphibians of India and
Sri Lanka (Checklist and Bibliography). Odys-
sey Publishing House, Bhubaneswar. (3) +
Mii 4 342 XX

Frost, D. R. (ed.). 1985. Amphibian Species of
the World: A Taxonomic and Geographical
Reference. Allen Press, Inc. and Association
of Systematics Collections, Lawrence. v +
132 Ds

GORHAM, S. W. 1974. Checklist of World Am-
phibians Up To January, 1970. The New
Brunswick Museum, St. John. 173 p.

INGER, R. F. AND S. K. DuTTA. 1986. An over-
view of the amphibian fauna of India. J.
Bombay Nat. Hist. Soc. 83 (Suppl.): 135-146.

LEVITON, A. E., R. H. GrBBs, JR., E. HEAL, AND
C.E. Dawson. 1985. Standards in herpetolo-
gy and ichthyology: Part I. Standard symbolic
codes for institutional resource collections in
herpetology and ichthyology. Copeia 1985:
802-832.

Manl, M.S. 1974. Physical features. p. 11-59.
In: Ecology and biogeography in India. M.S.
Mani (ed.). Dr. W. Junk Publishers, The
Hague.

WolLF, S. 1936. Revision der Untergattung Rha-
cophorus (ausschliesslich der Madagaskar-
Formen). Bull. Raffles Mus. 12: 137-217.

Accepted: 2 May 2000

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4
1

CURRENT HERPETOLOGY

MANAGING EDITOR
Masafumi MATSUI

Graduate School of Human and Environmental Studies, Kyoto University, Yoshida
Nihonmatsu-cho, Sakyo-ku, Kyoto 606-8501 Japan
(fumi@zoo.zool.kyoto-u.ac.jp)

ASSOCIATE EDITORS

Kraig ADLER, Department of Neurobiology and Behavior, Cornell University, Seeley G. Mudd
Hall, Ithaca, New York 14853-2702, USA (kka4@cornell.edu)

Aaron M. BAUER, Department of Biology, Villanova University, 800 Lancaster Avenue, Vil-
lanova, PA 19085 USA (aaron.bauer@villanova.edu)

Ilya S. DAREVSKY, Zoological Institute, Russian Academy of Sciences, St.Petersburg 199034
RUSSIA (Darevsky@herpet.zin.ras.spb.ru)

Indraneil DAS, Institute of Biodiversity and Environmental Conservation, Universiti Malaysia
Sarawak, 94300, Kota Samarahan, Sarawak, MALAYSIA (idas@mailhost.unimas.my)
Richard C. GORIS, Hatsuyama 1-7-13, Miyamae-ku, Kawasaki 216-0026 JAPAN (goris@

twics.com)

Ivan INEICH, Laboratoire des Reptiles et Amphibiens, Museum National d’Histoire Naturelle,
25 rue Cuvier 75005 Paris, FRANCE (ineich@cimrs1.mnhn.fr)

Ulrich JOGER, Hessisches Landesmuseum Darmstadt, Zoologische Abteilung, Friedensplatz 1,
D-64283 Darmstadt, GERMANY (u.joger@hlmd.tu-darmstadt.de)

Tamotsu KUSANO, Department of Biological Science, Graduate School of Science, Tokyo
Metropolitan University, Minami-ohsawa, Hachioji, Tokyo 192-0397 JAPAN (tamo@
comp.metro-u.ac.jp)

Colin McCARTHY, Department of Zoology, The Natural History Museum, Cromwell road,
London SW7 5BD, UK (cjm@nhm.ac.uk)

Hidetoshi OTA, Tropical Biosphere Research Center, University of the Ryukyus, Nishihara,
Okinawa, 903-0213 JAPAN (ota@sci.u-ryukyu.ac.jp)

Michihisa TORIBA, Japan snake Institute, Yabuzuka-honmachi, Nitta-gun, Gunma 379-2300

JAPAN (snake-a@sunfield.or.jp)

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FUTURE MEETING
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CONTENTS
Predation by the opossum Didelphis marsupialis on the rattlesnake Crotalus durissus
see aie Selma Maria Almeida-Santos, Marta Maria Antniazzi,

Osvaldo Augusto Sant’Anna, and Carlos Jared:::::: 1

Aphaniotis nasuta (de Jong, 1930), a junior synonym of A. ornata (Van Kidth de
Jeude, 1893) (Squamata: Agamidae)……" Hidetoshi Ota and Tsutomu Hikida:::::- 11

Conspecific and heterospecific pair-formation in Rana porosa brevipoda and Rana
nigromaculata, with reference to asymmetric hybridization -::::--:: Ryohei Shimoyama:::::- 15

Ant specialization in diet of narrow-mouthed toad, Microhyla ornata, from Amamioshima
Island of the Ryukyu Archipelago (ii Toshiaki Hirai and Masafumi Matsui:-:::- 27

Nomenclatural history and rediscovery of Rhacophorus lateralis Boulenger, 1883
(Amphibia: Rhacophoridae) SR eae Nat PS rer Ocha Miche Mo ORM io Suse, Sid Sao & Indraneil Das で CC ser 35

Instructions to contributors oe sae 0:5 «ce ove wie) oe «0/05, 0 » lai ule «ele sista) alsin stelle » efnlsietalselelelaiale aietat ae 41

ISSN 1345-5834

December 2000

。 FORMERLY THE JAPANESE JOURNAL OF HERPETOLOGY

Published by
上 比 HERPETOLOGICAL SOCIETY OF JAPAN
KYOTO

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THE HERPETOLOGICAL SOCIETY OF JAPAN

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Current Herpetology 19(2): 43-55., Dec. 2000
© 2000 by The Herpetological Society of Japan

Phylogenetic Position of Draco fimbriatus, with a Molecular
Perspective on the Historical Biogeography of the Genus

Draco (Reptilia: Agamidae)

MAsANAo HONDA|*, HipEetTosH1 OTA2, Snowrcgr SENGOKU3, AND

Tsutomu HIKIDA!

1 Department of Zoology, Graduate School of Science, Kyoto University, Sakyo, Kyoto

606-8502, JAPAN

2 Tropical Biosphere Research Center, University of the Ryukyus, Nishihara, Okinawa

903-0213, JAPAN

3 Japan Wildlife Research Center, Yushima, Bunkyo, Tokyo 113-0034, JAPAN

Abstract: The phylogenetic relationship of Draco fimbriatus with other con-
generic species was inferred from 848 base pairs of the mitochondrial 12S and
16S rRNA genes. The results confirmed that the closest affinity was between
D. fimbriatus and D. maculatus. Our results also suggested that D. lineatus
diverged first, followed by the D. cornutus—D. volans cluster, D. dussumieri,
the D. fimbriatus—D. maculatus cluster, and D. blanfordii in that order, leav-
ing D. obscurus and D. taeniopterus as monophyletic. The taxonomic diver-
sity of Draco in each area of Southeast Asia appears to have increased through

multiple colonizations rather than through in situ diversifications.

Key words:

INTRODUCTION

The genus Draco Linnaeus, 1758, consist-
ing of some 21 species, is one of the most
prominent genera of the family Agamidae,
characterized by the presence of a patagium
(a wing-like skin extension supported by
elongated ribs) along the flanks. This ge-
nus is distributed in southern India and
throughout Southeast Asia (Inger, 1983;
Musters, 1983; Lazell, 1987, 1992; Ross and
Lazell, 1990). Musters (1983), on the basis

* Corresponding author. Tel: +81-75-753-
4091; Fax: +81-75-753-4114.

E-mail address: panda@zoo.zool.kyoto-u.ac.jp.
(M. Honda)

Agamidae; Draco; Mitochondrial DNA; Phylogenetics

of cluster analysis of a distance matrix for
morphological characters, recognized two
major lineages within this genus that were
distnguished most simply by the direction of
the nostrils—outward or upward (Fig. 1).
Based on the DNA sequence data, Honda et
al. (1999b) negated this view, and recog-
nized at least four distinct lineages within
the genus. Their analyses, however, failed
to resolve the relationships of these major
lineages in detail.

Honda et al. (1999b) did not investigate
nine species of Draco. Of these, D. min-
danensis, D. spilopterus, and six recently
described or revalidated species (Lazell,
1987, 1992; Ross and Lazell, 1990) were
nevertheless surmised to be closely related

Fic. 1.

Current Herpetol. 19(2) 2000

D. blanfordii

D. obscurus

D. taeniopterus

D. melanopogon

D. haematopogon B
D. mindanensis

. Maximus

. quinquefasciatus

. dussumieri

D. fimbriatus
D. maculatus
D. lineatus

D. spilopterus
D

. volans

D. cornutus

Phylogenetic tree of the genus Draco, proposed by Musters (1983) on the basis of mor-

phological characters and geographic ranges. A: group of species characterized by outward-directed
nostrils and more or less developed nuchal crests; B: group of species characterized by upward-directed

nostrils and absence of distinct nuchal crests.

to species examined in their studies on the
basis of great morphological similarities.
The absence of those eight species from the
analyses does not thus seem to have im-
posed any crucial effect on the resultant
general picture of the phylogeny and bio-
geography of the genus (Honda et al.,
1999b). In contrast, absence of the other
species, D. fimbriatus, was supposedly more
problematic because its relationship with
other congeneric species was uncertain

(Musters, 1983).

In the present study, we sequenced a part
of mitochondrial DNA for D. fimbriatus
and analyzed the sequence data with com-
parable published data for other congeneric
species. Our purposes were to elucidate the
phylogenetic position of D. fimbriatus, and
to discuss the phylogenetic relationships
and historical biogeography of the genus
accordingly.

HONDA ET AL.—PHYLOGENY OF DRACO 45

MATERIALS AND METHODS

Samples Analyzed

A female Draco fimbriatus, collected
from Gunung Gate, West Java [Kyoto
University Zoological Collection (KUZ)
49655], was examined. We also incorpo-
rated into the analyses data published for
other Draco species, as well as for represen-
tatives of several other agamid genera
(Honda et al., 1999b, c, 2000) (Table 1).
Data for Bradypodion fischeri of the
Chamaeleonidae and Iguana iguana of the
Iguanidae, possible closest relatives of the
Agamidae and Acrodonta (i.e., an assem-

TABLE 1.

blage of the Chamaeleonidae and the
Agamidae), respectively (Frost and
Etheridge, 1989), were also incorporated
into the analyses (Ota et al., 1999; Honda et
al., 2000).

In some species of Draco, morphological
differentiations have been reported between
conspecific subspecies and/or populations
(e.g., Hennig, 1936; Taylor, 1963; Inger,
1983; Musters, 1983). However, con-
specific samples were designated as a single
operational taxonomic unit (OTU), because
our previous study showed that all con-
specific samples exclusively constitute
lowest clusters with high bootstrap values

Localities of Draco and other agamid samples used for analyses. Data sources are (a)

Honda et al. (1999a); (b) Honda et al. (1999b); (c) Honda et al. (2000); (d) Ota et al. (1999); (e) this

study. See Appendix for further details.

Sample Locality

D. blanfordii Thailand>

D. cornutus Borneo?

D. dussumieri India>

D. fimbriatus Java‘

D. haematopogon Peninsular Malaysia>
D. lineatus beccarii Sulawesi>

D. maculatus Thailand>

D. maximus Borneo?

D. melanopogon Thailand>

D. obscurus Borneo?

の . quinquefasciatus Peninsular Malaysia
D. taeniopterus Thailand>

D. volans Java?

Acanthosaura crucigera Thailand*

Agama 7677O

Aphaniotis fusca

Calotes versicolor
Gonocephalus grandis
Japalura polygonata
Lophognathus temporalis
Phoxophrys nigrilabris
Phrynocephalus axillaris
Physignathus cocincinus
Ptyctolaemus phuwuanensis
Bradypodion fischeri
Iguana iguana

West Asia or North Africa‘
Peninsular Malaysia>
Thailand‘

Peninsular Malaysia‘
Japan‘

New Guinea‘
Borneo‘

Central Asia‘
Thailand‘

Thailand>

Africa‘

America‘?

(Honda et al., 1999b).

The specific arrangement of Draco fol-
lows that by Musters (1983) (also see Honda
et al., 1999b). As in our previous study
(Honda et al., 1999b), we were unable to
examine D. mindanensis, D. spilopterus,
and six recently described or revalidated
species (Lazell, 1987, 1992; Ross and Lazell,
1990). Of these, D. biaro, D. bimaculatus,
and D. caerulhians were assumed to be
closest to D. lineatus, D. ornatus to D.
spilopterus, and D. everetti and D. jareckii
to D. volans on the basis of their great
morphological similarities. We _ believe
these designations do not lead to any sub-
stantial error in the results of the
phylogenetic analyses (Honda et al., 1999b).

Extraction, Amplification and Sequencing
of DNA

DNA extraction, amplification and se-
quencing are described in detail elsewhere
(Honda et al., 1999b, c). Approximately
850 base pairs (bp) of the mitochondrial 12S
and 16S rRNA genes were amplified using
the polymerase chain reaction (PCR; Saiki
et al., 1988) with primers L1091 and H1478
(Kocher et al., 1989), and L2606 and H3056
(Hedges et al., 1993), respectively.

Phylogenetic Analyses

Alignments for DNA sequences were de-
termined based on maximum nucleotide
similarity using CLUSTAL W 1.4 (Thomp-
son et al., 1994). The neighbor-joining
(NJ) method (Saitou and Nei, 1987) was ap-
plied to infer relationships among taxa with
a pairwise matrix of distance using
Kimura’s two-parameter model (Kimura,
1980). NJ analysis was performed with
PHYLIP 3.54c (Felsenstein, 1993).
Degrees of support for internal branches in
each tree were assessed by 1,000 bootstrap
pseudoreplications (Felsenstein, 1985). For
maximum-likelihood (ML) analysis and max-
imum-parsimony (MP) analysis, fast DNAml
1.0.6 (Olsen et al., 1994) and PAUP* 4.0b
(Swofford, 1998) with the heuristic search

Current Herpetol. 19(2) 2000

option were used, respectively. Confidence
was assessed by 1,000 bootstrap replica-
tions. In these three analyses, insertions
and deletions were excluded, and tran-
sition : transversion bias was assumed as
2: 1 according to the observed ratio for the
ingroup (transition/transversion = 1.96).

RESULTS

Aligned sequences from two mitochon-
drial genes (848 bp in total) are presented in
Fig. 2. The 12S rRNA fragment consisted
of a total of 412 sites, 303 of which were
variable. For the 16S rRNA fragment,
there was a total of 436 aligned sites, 260 of
which were variable. Inter-generic nucleo-
tide replacements within the Agamidae va-
ried from 133 bp (Calotes vs. Japalura) to
236 bp (Calotes vs. Lophognathus). Inter-
specific nucleotide replacements within Dra-
co varied from 48 bp (D. obscurus vs. D.
taeniopterus) to 120 bp (D. taeniopterus vs.
D. volans). Nucleotide replacements be-
tween D. fimbriatus and other congeneric
species were observed from 73 bp (vs. D.
maculatus) to 116 bp (vs. D. lineatus).

The NJ dendrogram derived from aligned
sequences is shown in Fig. 3A. Monophyly
of Draco was supported with complete
bootstrap proportions (BPs) (node 1:
BP=100%). Within the Draco cluster, we
recognized eight nodes (nodes 2-9), as more
or less substantial, because these were sup-
ported with BPs > 50%. Draco lineatus
diverged first, followed by the D. cornu-
tus—D. volans cluster (node 3: BP=99%),
の . dussumieri, the の . fimbriatus—D.
maculatus cluster (node 6: BP=97%), and
D. blanfordii in that order, leaving D. hae-
matopogon, D.maximus, D. melanopogon,
D. obscurus, D. quinquefasciatus, and D.
taeniopterus aS monophyletic (node 8:
BP=61%). Within node 8, D. obscurus
and D. taeniopterus formed an exclusive
cluster (node 9: BP=100%).

The results of ML (Fig. 3B) and MP ana-
lyses (Fig. 3C) showed no inconsistency

HONDA ET AL.—PHYLOGENY OF DRACO

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Current Herpetol. 19(2) 2000

Iguana iguana

Bradypodion fischeri

Agama stelio

99 Phrynocephalus axillaris
5 Lophognaths temparalis
100 Physignathus cocincinus

100

99 94 65

Ptyctolaemus phuwuanensis

Aphaniotis fusca
Calotes versicolor
Japalura polygonata
Acanthosaura crucigera

Gonocephalus grandis

51
Phoxophrys nigrilabris
D. lineatus
1 3 D. cornutus
100 99 D. volans
2 D. dussumieri
4 6 D. fimbriatus
79 97 D. maculatus
5
D. blanfordii
94
D. maximus
7
70 9 D. obscurus
8 100 D. taeniopterus
61

0.1

Iguana iguana
Bradypodion fischeri
Physignathus cocincinus

Lophognaths temparalis

D. melanopogon

D. quinquefasciatus

D. haematopogon

Phrynocephalus axillaris 50
Agama stelio
Ptyctolaemus phuwuanensis
Aphaniotis fusca 69
Phoxophrys nigrilabris
Gonocephalus grandis 95 94
Acanthosaura crucigera 60
Japalura polygonata
Calotes versicolor 67
D. lineatus
1 3 D. volans 1
D. cornutus 99
2 り . dussumieri 2
4 6 D. maculatus 94 4
D. fimbriatus 65
2 D. blanfordii 5
7 D. quinquefasciatus 78
8 D. haematopogon 7
72
D. melanopogon
D. maximus
9 D. taeniopterus
D. obscurus

Iguana iguana
Bradypodion fischeri
Agama stelio
Phrynocephalus axillaris
Lophognaths temparalis
Physignathus cocincinus
Ptyctolaemus phuwuanensis
Aphaniotis fusca
Gonocephalus grandis
Phoxophrys nigrilabris
Acanthosaura crucigera
Calotes versicolor
Japalura polygonata

D. lineatus

D. cornutus

D. volans

D. dussumieri

D. fimbriatus

D. maculatus

D. maximus

D. quinquefasciatus

D. blanfordii

D. melanopogon

D. haematopogon

D. obscurus

100 D. taeniopterus

HONDA ET AL.—PHYLOGENY OF DRACO 51

with the NJ dendrogram in terms of
branching topology of nodes 1-9 at the level
of BPs > 50% or P-values < 0.01 except
for the absence of node 8 in MP.

DISCUSSION

Phylogenetic relationships of Draco

Musters (1983) recognized two major
lineages within Draco (Fig. 1), that were
characterized by outward-directed nostrils
and more or less developed nuchal crests
(Group A), and upward-directed nostrils
and the absence of distinct nuchal crests
(Group B). Character states in the former
are reasonably assumed to be primitive
(Honda et al., 1999b). Musters (1983)
postulated the most basal divergence of D.
fimbriatus in Group A (Fig. 1).

Honda et al. (1999b), on the basis of
mitochondrial DNA sequence data, negated
this dichotomy because of the presence of
four distinct, deeply diverged evolutionary
lineages: D. lineatus, the D. cornutus—vo-
lans clade, D. dussumieri, and the clade
consisting of D. blanfordii, の . hae-
matopogon, D. maculatus, D. maximus, D.
melanopogon, D. obscurus, D. quinquefas-
ciatus and D. taeniopterus (henceforth
referred to as clade a). The relationship
resulting from the present analyses by in-
corporating the DNA sequence data from
D. fimbriatus also substantially differs from
that resulting from the morphological anal-
ysis by Musters (1983) in indicating the sis-
ter relationship of D. fimbriatus with D.
maculatus. This suggests that D. fimbria-
tus is a member of clade a, and that the

character states found in this species but
lacking in most other species including D.
maculatus (e.g., long head: Musters, 1983)
have evolved from states of corresponding
characters in D. maculatus.

The addition of D. fimbriatus to the anal-
ysis otherwise yielded results largely similar
to those of Honda et al. (1999b), and fur-
ther confirmed the close affinity between D.
dussumieri and node 5 (= clade a) among
the four major clades. The present results
also indicate that D. lineatus diverged first,
followed by the D. cornutus—D. volans
cluster and D. dussumieri in that order,
leaving node 5 as monophyletic.

With respect to the relationships of D.
blanfordii, D. obscurus and D. taeniopte-
rus, the present results, which point to the
non-monophyly and the sister relationship
between D. obscurus and D. taeniopterus,
contradict Inger’s (1983) and Musters’
(1983) views, which assumed the closest
affinity of these three species and their
relationships to be expressed as (D. taeniop-
terus, (D. blanfordii, D. obscurus)), respec-
tively. Moreover, our results suggest the
early divergence of D. blanfordii from the
other Southeast Asian species with derived
character states (see above). Morphologi-
cal similarities among these species (nuchal
fold, enlarged scales on gular pouch, and
five ribs in the patagium: Inger, 1983;
Musters, 1983) thus seem to be attributable
to convergence rather than to recent com-
mon ancestry. The character state of ob-
liquely upward directed nostrils (Inger,
1983, but described as “upward or slightly
posteriorly directed” by Musters [1983]) ob-

Fic. 3.

(A) NJ dendrogram of 26 taxa derived from a distance matrix of 12S and 16S rRNA sequence

data. Numbers beneath branches are bootstrap proportions (BPs) of at least 50% of the 1,000 boot-
strap pseudoreplications. Nodes with bold numbers are identical with ML and MP analyses. Bar
equals 0.1 unit of Kimura’s two-parameter distance. (B) ML dendrogram (In likelihood =-10,519.6).
All branches were supported as significantly positive (p く 0.01). Bar equals 0.1 unit. (C) MP clado-
gram using the heuristic search option (3,015 steps, 409 bp informative under the condition of par-
simony, consistency index=0.408). Branches without BP values were not supported in > 50% of the
replicates.

52 Current Herpetol. 19(2) 2000

Fic. 4. A map of Southeast and South Asia showing distributions of Draco species. Five areas are
recognized on the basis of distributional patterns in Draco and other taxonomic groups (Wallace 1860;
Dunn and Dunn, 1977; Musters, 1983; Heaney, 1984, 1991; Alcala, 1986; Holloway, 1987; Lazell,
1987, 1992; Ross and Lazell, 1990) following Honda et al. (1999b): 1, southern India; 2, Indochinese
Peninsula; 3, Malay Peninsula and the Greater Sunda Islands exclusive of Sulawesi; 4, Lesser Sunda
Islands, Sulawesi and Maluk Islands; 5, Philippines. Draco dussumieri is the only species distributed
in Area 1, D. blanfordii, D. maculatus, and D. taeniopterus in Area 2, D. cornutus, D. fimbriatus, D.
haematopogon, D. maximus, D. melanopogon, D. obscurus, and D. quinquefasciatus in Area 3, D.
lineatus in Area 4, and D. mindanensis and D. spilopterus in Area 5. Of these, D. blanfordii, D.
maculatus, and D. taeniopterus also occur in the northern part of Area 3, whereas D. obscurus is dis-
tributed in the southern part of Area 2 as well across the boundary in the Malay Peninsula. Draco
lineatus, while mainly occuring in Area 4, also occurs in the southeastern part of Area 3 and the
southern part of Area 5 as well. Draco volans has the widest distribution which, while seemingly
centering in Area 3, also partially encompasses Areas 2, 4 and 5.

HONDA ET AL.—PHYLOGENY OF DRACO 53

served in D. blanfordii, may represent an
intermediate state between the two extreme
states of this character (outward and up-
ward directions: see above).

Biogeography of Draco

Figure 4 depicts the phylogeographical
hypothesis of Draco resulting from the
present analyses. Honda et al. (1999b)
divided the geographic range of Draco into
five areas (as numbered in Fig. 4), and con-
structed the zoogeographical scenario con-
sisting of five stages as follows: (i) The
presumptive ancestor of Draco first possibly
diverged into three groups, the ancestors of
の . lineatus, the D. volans—cornutus
cluster, and the cluster consisting of D. dus-
sumieri and clade a (see above) through a
series of vicariances among Areas 4, 3, and
2; (ii) the common ancestor of D. dus-
sumieri and clade a invaded Area 1 from
Area 2, and there (area 1) it was isolated and
diverged into D. dussumieri; (iii) the com-
mon ancestor of clade a invaded Area 3
from Area 2, and there (Area 3) the
Southeast Asian members of Group B
diverged from the maculatus-like ancestor;
(iv) the resultant ancestors of D. blanfordii
dispersed into Area 2 from Area 3; and (v)
invasion of Area 2 by the ancestor of D.
taeniopterus from Area 3 followed (iv). Of
these, however, the reliability of stages (i),
(ii), and (ili) were not supported with
sufficient BPs in the DNA sequence or allo-
zyme analysis by Honda et al. (1999b).

The present results support these five
hypothetical stages with significant BPs or
P-values, enabling us to extend the scenario
for the historical biogeography of Draco as
follows. (I) The primary divergence of
Draco occurred between D. lineatus in Area
4 and the other (node 2) occurred in Areas 2
and 3. (II) The common ancestor of the
latter (node 2) further split into the D.
volans—cornutus clade (node 3) in Area 3
and the other (node 4) split in Area 2. (IID)
The ancestor of D. dussumieri diverged in
Area 1 from the common ancestor of node

4, leaving node 5 (=clade a: Honda et al.,
1999b) as monophyletic. (IV) The common
ancestor of node 5 (=clade a), originally
distributed in Area 2, dispersed into Area 3,
and then it split into the D. fimbriatus—
maculatus clade (node 6) and the Southeast
Asian members of Group B (node 7) in
Areas 2 and 3, respectively. (V) The ances-
tor of node 7 further diverged into several
species in Area 3. (VI) The ancestors of D.
fimbriatus independently dispersed from
Area 2 into Area 3. (VII) The ancestors of
D. blanfordii invaded Area 2 from Area 3.
(VIII) The ancestor of D. taeniopterus in-
dependently dispersed from Area 3 into
Area 2. Hypothetical stages (VII) and
(VIII) are consistent with stages (iv) and (v)
of Honda et al. (1999b), respectively.
These two stages seem to be supported by
the early divergence of D. blanfordii from
node 8, and by the sister relationship be-
tween D. taeniopterus and D. obscurus
(node 9) in the present study. This idea is
also circumstantially supported by the fact
that D. blanfordii has diverged into several
subspecies within Area 2, whereas D.
taeniopterus is rather monomorphic
(Musters, 1983), and that D. blanfordii dis-
persed into the northern and eastern parts
of Area 2, whereas D. taeniopterus is
confined to the southern part of this region
(Taylor, 1963; Musters, 1983).

Two endemic species, D. mindanensis
and D. spilopterus, are distributed in Area
5. Diversity in Area 5 is assumed to have
increased through multiple colonizations
from outside (D. mindanensis from Area 3,
and D. spilopterus from Area 3 or Area 4),
not through an endemic radiation (Honda
et al., 1999b). In Area 4 where the range of
D. lineatus is centered, D. volans also oc-
curs. Occurrence of D. volans in this area
seems to have resulted from a relatively re-
cent dispersal from Area 3. The taxonomic
diversity of Draco in Areas 2 and 3 is also
considered to have increased through multi-
ple colonizations. We suspect that the
diversity of this genus in each area of the

Current Herpetol. 19(2) 2000

Southeast Asian region has increased chiefly
through multiple colonizations rather than
through in situ diversifications.

ACKNOELEDGMENTS

We would like to thank M. Kobayashi for
laboratory assistance and K. Araya for
providing specimens of D. fimbriatus. We
are also much indebted to N. Murakami for
useful suggestions on phylogenetic analyses.
Special thanks are due to N. Satoh and
members of his laboratory for continuous
support for our laboratory experiments.
Experiments were also carried out using the
facilities of the Kyoto University Museum.
Our research was partially supported by the
Nakayama Foundation for Human Science.

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APPENDIX

Accession Numbers of Specimens.—The
DDBJ accession numbers of 12S and 16S rRNAs
are presented.

Acanthosaura crucigera: 4B031963, AB031980.
Agama stelio: AB031976, AB031993. Aphaniotis
fusca: AB023749, AB023771. Calotes versicolor:
AB031964, AB031981. Draco blanfordii blanfor-
dii: AB023733, AB023751. D. cornutus: AB023728,
AB023752. D. dussumieri: AB023734, AB023753.
D. haematopogon: AB023735, AB023754. D. linea-
tus beccarii: AB023737, AB023756. D. macula-
tus maculatus: AB023739, AB023758. D. maxi-
mus: AB023740, AB023760. D. melanopogon:
AB023741, AB023761. D. melanopogon: AB023742,
AB023762. D. obscurus obscurus: AB023743, AB
023763. D. quinquefasciatus: AB023745, AB023765.
D. taeniopterus AB023747, AB023767. D. volans
volans: AB023748, AB023770. Gonocephalus
grandis: AB031966, AB031983. Japalura poly-
gonata polygonata: AB031968, AB031985. Phoxo-
Dhrys nigrilabris: ABO031971, AB031988. Phrynoce-
phalus axillaris: ABO31972, AB031989. Physig-
nathus cocincinus: AB031973, AB031990. Pty-
ctolaemus phuwuanensis: AB023750, AB023772.
Bradypodion fischeri: AB031962, ABO31979.
Iguana iguana: AB028742, AB028756.

Accepted: 7 April 2000

ea,

iva

Current Herpetology 19(2): 57-61., Dec. 2000
© 2000 by The Herpetological Society of Japan

Mabuya cumingi (Reptilia: Scincidae): An Addition to the

Herpetofauna of Lanyu Island, Taiwan

HIDETOSHI OTA!* AND WEN-SAN HUANG?

! Tropical Biosphere Research Center, University of the Ryukyus, Nishihara, Okinawa

903-0213, JAPAN

< Zoological Department, National Museum of Natural Science, 1, Kuan Chien Road,

Taichung, 404 Taiwan, REPUBLIC OF CHINA

Abstract: An adult male Mabuya, recently collected from Lanyu Island, Tai-
wan, was identified as Mabuya cumingi, a species hitherto known only from
Luzon Island of the Philippines, on the basis of small body size (56.6 mm in
snout-vent length), embossed dorsal head scales, five scales beneath toe I, and
the presence of a dark middorsal stripe. Occurrence of this species on Lanyu
Island was also confirmed by additional sighting records. Mabuya cumingi is
regarded as a fourth reptile species representing a dispersal to this island from

the Philippines.

Key words:
land; Taiwan; Philippines

INTRODUCTION

Lanyu Island is an islet of 45.7 km? in
area and 548 m in height, and is located ca.
60 km southeast of the main island of Tai-
wan and ca. 390 km north of Luzon Island,
the Philippines. Since the description of
Gekko kikuchii by Oshima (1912), a total of
18 species of reptiles, including three possi-
ble human commensals (Hemidactylus
frenatus, Hemiphyllodactylus typus typus,
and Ramphotyphlops braminus) and 15
putative native species, have been recorded
from this island (Ota, 1991a,b: Lue et al.,
1999).

A number of previous biogeographers ar-

* Corresponding author. Tel: +81-98-895-
8937; Fax: +81-98-895-8966.

E-mail address: ota@sci.u-ryukyu.ac.jp (H.
Ota)

Mabuya cumingi; Mabuya multicarinata; Scincidae; Lanyu Is-

gued that the fauna of Lanyu Island is
characterized by a higher propotion of
Philippine elements when compared with
that of the main island of Taiwan (e.g.,
Kuroda, 1925; Kano, 1933, 1936). Of the
15 putative native reptiles hitherto known
from Lanyu Island, three (G. kikuchii,
Lepidodactylus yami, and Mabuya mul-
ticarinata borealis) are classified as the
Philippine elements, whereas most of the
remaining species are considered to
represent dispersals from the main island of
Taiwan (Ota, 1991a,b).

Recently, one of us (WH) collected a
specimen of a strange skink from Lanyu Is-
land. This specimen, deposited in the her-
petological collection of the National Muse-
um of Natural Science, Taichung, as NUNS
3371, had keeled scales on the dorsum of
the body and completely separated
pterygoids with a palatal notch extending

forwards to the level of the center of the
eye, and thus was identified as a member of
the genus Mabuya (see Greer [1970, 1977]).
In this paper, we describe external charac-
ters of this specimen in detail, and compare
it with species of Mabuya known from Tai-
wan and the Philippines to determine its
specific status.

MATERIAL AND METHODS

The specimen was collected by hand from
sparse grass near the coast, 3 km north of
Tung-Ching Village, Lanyu Island
(22°04'N, 121°33’E), on 17 July 1999. Af-
ter being photographed in life (Fig. 1), the
lizard was fixed in 10% formalin, perserved
in 70% ethanol, and then subjected to
detailed examination of morphological
characters. Measurements were taken to
the nearest 0.1mm with dial calipers.
Definitions of characters follow those of
Brown and Alcala (1980).

RESULTS

Description of the present specimen
A male with well-developed testes and

Current Herpetol. 19(2) 2000

convoluted epididymides; snout-vent length
(SVL) 56.6 mm; tail length 99.4 mm; head
length 12.2 mm; head width 8.1 mm; snout-
eye length 4.7 mm; eye length 3.5 mm; ear
length 1.1mm; snout-forelimb distance
20.8mm; axilla-groin distance 28.8 mm;
forelimb length 17.7 mm; hindlimb length
25.8 mm.

Snout largely tapered, but rounded at tip;
scales on dorsal surface of head embossed;
rostral approximately twice as broad as
high, rounded dorsally, in contact with
frontonasal; supranasals long, narrow, not
in contact at midline; frontonasal nearly as
broad as frontal, in contact with frontal;
prefrontals separated at midline; frontal
elongate, broadly in contact with second
supraoculars; supraoculars four, first
smallest, second largest; frontoparietals not
fused; interparietal incomplete, fused to
parietals posteriorly; nuchals in two pairs;
postnasal absent; anterior loreal distinctly
higher than long, much shorter than
posterior loreal; supralabials seven, fifth
largest, beneath eye; seven infralabials; ear
small, without lobules; each scale on dorsal
and lateral surfaces of body with six or
seven keels, ventral scales smooth, or slight-

i, ae a WW 0
07 i 4 cd

Fic. 1. Dorsolateral view of the specimen of Mabuya cumingi from Lanyu Island, Taiwan (NMNS

3371: SVL=56.6 mm).

OTA & HUANG—LIZARD FROM TAIWAN

ly rugose; 30rows of scales around mid-
body; 43 vertebral scale rows between
parietals and base of tail just above vent;
preanals slightly enlarged; limbs well deve-
loped; finger III almost as long as finger IV;
toe IV distinctly longer than toe III; subdig-
ital scales 5-5, 9-9, 14-14, 15-15 and -8 on
left-right fingers I, II, III, IV and V (left
finger V damaged), and 5-5, 10-10, 15-16,
21-21 and 13-13 on left-right toes I, II, III,
IV and V, respectively; tail not damaged or
regenerated.

In life, dorsal surfaces of head and body
light grayish tan, with black middorsal
stripe; additional longitudinal black stripes,
separated from each other by white inter-
spaces, in dorsolateral, lateral, and ven-
trolateral regions; lateral stripe, running
from nasal and eye to above base of hin-
dlimb broad, the others narrower (Fig. 1).
In preservative, dorsal ground color and
white interspaces faded to dark gray and
light gray, respectively, making longitudinal
dark stripes indistinct.

Comparisons

Three species of Mabuya, M. longicauda-
ta, M. multifasciata, and M. multicarinata
borealis, have been recorded from Taiwan
(Ota et al., 1994; Lue et al., 1999). The
present specimen differs from the three spe-

TABLE 1.
M. indeprensa from the Philippines.

cies in a much smaller body size, because
adult SVLs of these species well exceed
60mm (Okada et al., 1992; Ota et al.,
1994). Furthermore, it differs from M. /on-
gicaudata and M. multifasciata in having
distinctly embossed head scales, because in
the latter species, scales covering the dorsal
surface of head are smooth, or only slightly
rugose (Ota et al., 1994). From M. mul-
ticarinata borealis, the present specimen
also differs in having a greater number of
vertebral scale rows (43, vs. 34-41), fewer
scales beneath toe IV (21, vs. 24-28), and a
distinct middorsal stripe (vs. no middorsal
stripe: Ota, 1991a).

Most of those and other characteristics of
NMNS 3371 are shared with M. cumingi
and M. indeprensa from the Philippines
(Brown and Alcala, 1980). So, we compare
the present specimen with these two species
in detail (Table 1). Brown and Alcala
(1980) reported that both of these Philip-
pine species have a distinct interparietal
which almost always separates parietals
completely. In the present specimen,
however, the interparietal is incomplete be-
cause it is fused with parietals posteriorly.
The Lanyu specimen differs from M. in-
deprensa in having shorter hindlimbs in re-
lation to the axilla-groin distance, and fewer
scales beneath toe I. States of these and

Comparisons of characters among the Lanyu specimen of Mabuya, and M. cumingi and
Abbreviations are: SVL, snout-vent length; HLL, hindlimb

length, AGD, axilla-groin distance; IP, interparietal; MSR, midbody scale rows; VS, vertebral scale
rows; TIS, subdigital scales beneath the toe I; TIVS, subdigital scales beneath the toe IV.

M. cumingi* M. indeprensa*

Character Lanyu specimen
SVL (mm) 56.6
HLL/AGD (%) 90.0
HLL/SVL (%) 45.6

iP incomplete
MSR 30

VS 43

TIS 5)

TIVS |

39.5-54.0 45 .6-66.6
74-96 94-116
37-45 45-58

distinct distinct
28-32 30-34
40-47 41-48
5-6 6-8
16-21 18-24

* Data for these species in the Philippines were taken from Brown and Alcala (1980).

most other characters in the present speci-
men are shared with M. cumingi, except for
SVL, which is slightly greater in the Lanyu
specimen than in the latter.

DISCUSSION

Morphological comparisons indicate that
the Lanyu specimen most resembles M.
cumingi, but differs from the species in hav-
ing slightly greater SVL and an incomplete
interparietal. Considering that the body
size often varies extensively among con-
specific insular populations in reptiles (see
Ota et al [1995] and papers referred therein
for examples), such a difference in SVL be-
tween the present specimen and M. cumingi
from the Philippines may have little tax-
onomic significance. Moreover, it is proba-
ble that the interparietal condition in the
Lanyu specimen represents an anomaly
rather than a stable population feature.
We thus identify the specimen as M.
cumingi. Further detailed analysis of vari-
ation on the basis of additional specimens is
desired to verify those assumptions.

During the fieldwork on Lanyu Island in
1995 (by HO) and 1997-1999 (by WH), both
of us observed several other individuals
with body size and color pattern similar to
those of the present specimen. Thus,
although the present record was made only
on the basis of a single voucher specimen,
we are almost certain that there is an estab-
lished breeding population of M. cumingi
on Lanyu Island. All individuals sighted
were found in relatively open environments
near the coast, and appeared to be most ac-
tive on clear mornings and at dusk.

In the synopsis of the lizards of Taiwan,
Lin and Cheng (1990) provided three color
photographs as those of M. multicarinata.
The animals photographed, however,
showed a color pattern almost identical with
that of the present specimen including a dis-
tinct middorsal stripe on body. Therefore,
although the locality of the individual pho-
tographed was not given in that book, it is

Current Herpetol. 19(2) 2000

most likely that it actually represented M.
cumingi from Lanyu Island rather than M.
multicarinata.

Since the original description by Brown
and Alcala (1980), Mabuya cumingi has
been known only from Luzon Island.
Thus, the present finding seems to represent
another example of reptilian dispersals to
Lanyu Island from the Philippines (Ota,
1991a, b).

ACKNOWLEDGMENTS

HO thanks M. Toda and S. Iwanaga for
helping with his fieldwork on Lanyu Island,
and WH is grateful to C. H. Chang for his
assistance. This research was partially sup-
ported by grants from Kuo Wu Hsiu Luan
Foundation and National Museum of
Natural Science (to WH), and from the
Fujiwara Natural History Foundation (to
HO).

LITERATURE CITED

Brown, W.C. AND A.C. ALCALA. 1980.
Philippine Lizards of the Family Scincidae.
Silliman University Press, Dumaguete, Philip-
pines. i-xi+ 264 p.

GREER, A. E. 1970. A subfamilial classification
of scincid lizards. Bull. Mus. Comp. Zool.
139(3): 151-184.

GREER, A. E. 1977. The systematics and evolu-
tionary relationships of the scincid lizard ge-
nus Lygosoma. J. Nat. Hist. 11(3): 515-540.

Kano, T. 1933. Biogeographical studies of
Kotosho. 2. Geol. Rev. Japan 9(6): 475-491.
(in Japanese)

Kano, T. 1936. Some problems concerning the
biogeography of Kotosho, near Formosa. 7.
Geol. Rev. Japan 12(12): 1107-1133. (Gn
Japanese with English abstract)

Kuropa, N. 1925. On the distribution of
Japanese birds. Chigaku Zasshi 37: 369-380.
(in Japanese)

Lin, J.-Y. AND H.-Y. CHENG. 1990. A Synop-
sis of the Lizards of Taiwan. Taiwan Museum,
Taipei. i-xiii+ 176 p. (in Chinese)

Lug, K.-Y., M.-C. Tu, AND G. SHANG. 1999. A
Pictorial Guide to the Amphibians and Rep-
tiles of Taiwan. Union for the Nature Conser-

OTA & HUANG—LIZARD FROM TAIWAN 61

vation, ROC & Great Nature Press, Taipei,
Taiwan. (in Chinese)

OkApA, S., H. Ota, M. HAsEGAwa, T. HIKTDA,
H. Mryacuni, AND J. KAro. 1992.
Reproductive traits of seven species of lygoso-
mine skinks (Squamata: Reptilia) from East
Asia. Nat. Hist. Res. 2(1): 43-52.

OsgrwA, M. 1912. Description of a new gecko
from Botel Tobago Island. Philippine J. Sci.
7(4): 241-242, pls.1-2.

Ota, H. 1991a. Taxonomic status of Mabuya
multicarinata (Gray, 1845)(Scincidae:
Squamata: Reptilia) from Taiwan, with com-
ments on the herpetofauna of Lanyu Island.
Bull. Coll. Sci., Univ. Ryukyus (51): 11-18.

Ota, H. 1991b. Systematics and biogeography
of terrestrial reptiles of Taiwan. p. 47-112. In:

Y.-S. Lin and K.-S. Chang (eds.), Systematics
and Biogeography of Terrestrial Reptiles of
Taiwan. Council of Agriculture, Taipei, Tai-
wan.

Ota, H., H.-W. CHANG。 K.-C. Liv, AND T.
HrIkrpA. 1994. A new record of the
viviparous skink, Mabuya _ multifasciata
(Kuhl, 1820)(Squamata: Reptilia), from Tai-
wan. Zool. Stud. 33(1): 86-89.

Ota, H., M. SgrRoMA, AND T. Hixipa. 1995.
Geographic variation in the endemic Ryukyu
green snake Cyclophiops semicarinatus (Ser-
pentes: Colubridae). J. Herpetol. 29(1): 44-50.

Accepted: 7 April 2000

Current Herpetology 19(2): 63-70., Dec. 2000
© 2000 by The Herpetological Society of Japan

Sperm Morphology of Some Indian Frogs as Revealed

by SEM

Mitsuru KURAMOTO ANp S. HAREESH JOSHY2

1 3-6-15 Hikarigaoka, Munakata, Fukuoka 811-3403, JAPAN

2 Department of Zoology, St. Aloysius College, Mangalore-575 007, INDIA

Abstract: The size and shape of 15 species of frogs (Ranidae and
Rhacophoridae) from southwestern India were investigated by light and scan-
ning electron microscopy. Most species of the genus Rana had spermatozoa of
a typical form, with a thick sperm head and a thin tail. In contrast, Rana
beddomii had long spermatozoa with a slender and densely coiled sperm head
and a thick tail, suggesting the validity of the genus Indirana. The sperm head
of Nyctibatrachus major was thick and very loosely coiled. Differing from
Rhacophorus species from east Asia, the sperm head of Rh. malabaricus was
not coiled. Polypedates maculatus had very long thread-like spermatozoa as
in Rh. malabaricus. Spermatozoa of all examined species of the genus
Philautus had a crescent-like sperm head and a thin tail resembling the head

and tail of the genus Chirixalus.

Key words:

INTRODUCTION

To date, spermatozoa of more than a
hundred species of frogs have been investi-
gated by light microscopy (LM), SEM or
TEM (e.g., Lee and Jamieson, 1992;
Kuramoto, 1998; van der Horst et al.,
1995). From these studies, it became evi-
dent that sperm morphology is very variable
between taxa and useful for elucidating tax-
onomic relationships between them. For
example, Kuramoto (1996) showed that
four rhacophorid genera from Japan and
Taiwan have each a distinctive form of
spermatozoa and, within the genus Buerger-
ja, the form of the spermatozoon of B.
buergeri is apparently derived from the

* Corresponding author. Tel: 十 81-940-32-
0730

Anura; Spermatozoa; SEM; India

sperm form of its congeners. It was also
suggested that sperm morphology reflects
the mode of reproduction (van der Horst et
al., 1995).

India has a rich amphibian fauna, involv-
ing more than 180 species (Inger and Dutta,
1986). Among these, only two species,
Bufo stomaticus and Rana tigerina, have
been studied spermatologically (Sharma
and Dhindsa, 1955; Sharma and Sekhri,
1955). The primary purpose of these stu-
dies was to clarify the process of spermato-
genesis and thus detailed descriptions of
sperm are not given.

We examined sperm morphology of
several frog species from Karnataka and
Kerala, southwestern India by light
microscopy (LM) and scanning electron
microscopy (SEM). Dubois (1992) revived
several subfamilies and genera in the Rani-

dae and his revision has induced much con-
fusion and debate on their validity. In our
present materials representatives of some of
his genera are involved, and we expect that
our spermatological work will provide cues
to resolve this taxonomic problem. Also,
we examine generic differentiation in the
Rhacophoridae including the genera Philau-
tus for which sperm morphology has not yet
been reported.

MATERIALS AND METHODS

Frogs used in this study were: Rana tem-
poralis from Madikeri, Karnataka; R.
malabarica from Cannanore, Kerala; R.
limnocharis from Madikeri; R. keralensis
from Madikeri; R. syhadrensis from Man-
galore, Karnataka; R. cyanophlyctis from
Mangalore; R. tigerina from Mangalore; R.
beddomii from Madikeri; Nyctibatrachus
major from Madikeri (Ranidae);
Rhacophorus malabaricus from Madikeri;
Polypedates maculatus from Madikeri;
Philautus sp. A from Mangalore; Philautus
sp. B from Mangalore; Philautus sp. C
from Kudremukh, Karnataka; and Philau-
tus sp. D from Kudremukh (Rhacophori-
dae). All specimens were collected in June
and July 1999.

Philautus species, small arboreal frogs,
are difficult to identify as Inger and Stueb-
ing (1997, p. 163) admitted, and our speci-
mens did not seem to fit the descriptions of
Philautus species from the Western Ghats
(Ahl, 1931; Daniels, 1998; Inger et al., 1984;
Rao, 1937). Brief descriptions of our
specimens are given here: Philautus sp. A
was reddish brown on the back, some with
irregular dark blotches, underside immacu-
late, tympanum about half of eye diameter
and lower half of tympanum white, snout
rather pointed, ca. 24mm in SVL; Philau-
tus sp. B was pale gray on the back with an
inverted irregularly U-shaped blackish
mark, underside with many scattered small
dark markings, tympanum small and in-
distinct, snout not pointed, ca. 25mm in

Current Herpetol. 19(2) 2000

SVL; Philautus sp. C was dark brown on
the back with a large hourglass-like pale
marking from eye to vent, underside ver-
miculate with brown, yellow, and white,
two distinct large yellowish spots on an-
terior surface of thigh, snout not pointed,
ca. 18mm in SVL; Philautus sp. D was pale
yellow to reddish yellow on the back and
underside including the vocal sac, some
with three dark indistinct longitudinal
stripes on the back, tympanum small and
indistinct, ca. 26mm in SVL.

We collected several other unidentified
species in the genera Rana and Philautus,
and we will refer to sperm morphology of
these species briefly.

The testis was squashed in a small quanti-
ty of water with forceps, and the sperm sus-
pension thus prepared was put on slides and
on cover slips, fixed with 2% glutaraldehyde
for about one hour, and air-dried. Sper-
matozoa on the slide were stained with
Giemsa for LM and those on cover slips
were coated with gold and observed with a
scanning electron microscope JSM-T200
(JEOL). Two males were used for each
species.

RESULTS

Excepting Rana beddomii, all seven spe-
cies of the genus Rana had very similar
spermatozoa with a thick sperm head and a
thin tail (Fig. 1). Sperm sizes were also
similar, ranging from about 15 to 20 um in
head length and from about 45 to 70 ym in
total length, but variable in head width
(Table 1). The acrosome and the middle
piece or neck piece were discernible exter-
nally from the head proper by their slightly
narrower width. The tail was about 0.2 um
in width. Three unidentified species from
Mangalore and Kudremukh, apparently be-
longing to the Rana limnocharis complex,
had spermatozoa which were similar to
those of the above species.

Spermatozoa of R. beddomii were very
long, about twice as long as those of the

KURAMOTO & JOSHY—FROG SPERM MORHOLOGY

Fic. 1. Spermatozoa of Rana temporalis (A), R. malabarica (B), R. limnocharis (C), R. keralensis
(D), R. syhadrensis (E), R. cyanophlyctis (F), and R. tigerina (G). Scales equal 10 um.

gated, densely and sinistrally coiled spiral,
about 0.5m in diameter of spiral and

other species of the genus Rana (Fig. 2,
Table 1). The sperm head formed an elon-

TABLE 1. Sperm sizes in 15 species of frogs from southwestern India (Mean+SD in um, N=10).

Head length Tail length Total length Head width
Rana temporalis 20.3 +£0.82 45.4+3.26 (Haas 55745) L720. 10
Rana malabarica 17.21.49 27.9+2.78 45.0+ 2.81 1-62-0107
Rana limnocharis 17.4+1.28 tae sells S526 3:36 1.2+0.09
Rana keralensis 16.5+0.94 40.4+4.25 56.9+ 4.67 Loh 0206
Rana syhadrensis 18.3+2.90 367 ste ll 555025) 9216 0.9+0.07
Rana cyanophlyctis 17e20e=205 54.2+3.78 (Meas Tey) 0.9+0.05
Rana tigerina 15.0 土 1.43 47.1 土 2.99 6221 == 3.67 0.90.08
Rana beddomii 37.5+0.90 79.2 土 5.99 Kees Ge 0.5+0.03
Nyctibatrachus major 23 2O=t2 3/12 40.1+7.11 63.94 7.28 0.80.06
Rhacophorus malabaricus SS ana 92.6 土 6.93 144.9+ 5.96 0.6+0.05
Polypedates maculatus 76.5+4.20 84.0 土 14.1 160.5+14.6 0.4+0.05
Philautus sp. A 21.6+2.76 28.4 土 4.74 SOLQsE 5221 0.8+0.05
Philautus sp. B DS = ail 28.5+2.66 24322 4.38 0.80.04
Philautus sp. C 19.0+2.28 30.9+4.88 49.9+ 4.56 0.80.03
Philautus sp. D DOR ==3E59 29a] = 3250 S053 5.03 OF85=20205

Current Herpetol. 19(2) 2000

Fic. 2. Spermatozoa of Rana beddomii (A), Nyctibatrachus major (B), and a part of the coiled

sperm head of R. beddomii (C). The lower part of the coil in (C) is the basal portion of the head.

Scales equal 10 um in (A) and (B), and 1 ym in (C).

about 0.3 um in width of coil fiber. “Head
length” of R. beddomii in Table 1 shows the
length of the coil, not the net length of coil
fiber. The tail width (about 0.5 “m) was
nearly the same as head width and much
wider than those of its congeners. This
type of spermatozoa is new to our
knowledge of anuran sperm morphology.
We examined the sperm of a frog from
Madikeri which was similar to R. beddomii
but could not be precisely identified. This
frog had spermatozoa which were nearly
identical to those of R. beddomii. The tes-
tis of R. beddomii and the unidentified bed-
domii-like frog was considerably larger than
that of the other ranid frogs.
Nyctibatrachus major had also peculiar

spermatozoa (Fig. 2). Typically the sperm
head was S-shaped, undoubtedly forming a
loose coil three-dimentionally. A slightly
thin portion at the tip of head may be the
acrosome and that at the end may be the
middle piece. The tail was 0.13 wm in
width, much thinner than most species of
the genus Rana. Total sperm length was
within the range of most Rana species.
This type of spermatozoa has not been
reported in Ranidae. The testis was much
smaller than in most ranid species.
Rhacophorus malabaricus had _ long
thread-like spermatozoa, and Polypedates
maculatus had still longer ones (Fig. 3,
Table 1). The acrosome and the middle
piece could not be distnguished externally in

KURAMOTO & JOSHY—FROG SPERM MORHOLOGY 67

Fic. 3.
sp. A (D), Philautus sp. B (E), and Philautus sp. D (F). (A) and (C) were taken by LM. Scales equal
20 vm in (A) and (B), and 10 vm in (C)-(F).

either species. The tail was thick, only
slightly thinner than the sperm head with a
width of about 0.4 to 0.5 wm.

The four species of the genus Philautus
had spermatozoa of nearly the same size as
those in the genus Rana (Table 1). The
sperm head was invariably bent slightly,
forming a crescent (Fig. 3). The middle
piece was granular in appearance. The tail
was thin, about 0.2 ym in width. We exa-
mined sperm of three other species of
Philautus (unidentified) and all were very
similar to that of the above four species in
every characteristic.

DISCUSSION

Sperm shape and size of the seven species
of the genus Rana examined in this study
(temporalis, malabarica, limnocharis, ker-
alensis, syhadrensis, cyanophlyctis, tigeri-

Spermatozoa of Rhacophorus malabaricus (A, C), Polypedates maculatus (B), Philautus

na) are essentially identical to those of the
other Rana species reported so far, in that
all have a thick sperm head and a thin tail
(Kuramoto, 1998; van der Horst et al.,
1995). The sperm form of R. figerina
agrees well with the description by Sharma
and Sekhri (1955). The sperm head of R.
limnocharis is much longer than that of the
same species from Japan and China
(Kuramoto, 1998), suggesting intraspecific
geographic variations in sperm size as exem-
plified in Japanese salamanders (Kuramoto,
1997). Noticeable exceptions in the genus
Rana were three species of the Rana narina
complex and R. ishikawae which had very
long, slender spermatozoa (Kuramoto,
1998).

Rana beddomii had a completely different
type of sperm which was unlike any other
sperm type found in anurans. The sperm
head is long and densely coiled, and the tail

Current Herpetol. 19(2) 2000

is thick suggesting involvement of filamen-
tous components other than a single flagel-
lum. Detailed TEM analysis is needed to
clarify this unusual feature of sperm. In
the Rana species here examined, /im-
nocharis, keralensis, and syhadrensis are
frequently allocated to the genus Lim-
nonectes, cyanophlyctis to the genus Eu-
phlyctis, tigerina to the genus
Hoplobatrachus (as H. tigerinus), and bed-
domii to the genus Jndirana (Duellman,
1993; Dutta and Manamendra-Arachchi,
1996). The genera Limnonectes, Euphlyc-
tis, and Hoplobatrachus do not show any
sign of sperm differentiation as already
pointed out by Kuramoto (1998) in Lim-
nonectes and Hoplobatrachus, whereas In-
dirana differs completely from other genera
in sperm type, indicating a different phyletic
lineage. The karyotype of R. beddomii
(2n= 24, Joshy et al., 1999) also differs from
the other Indian ranid frogs. These suggest
strongly the validity of the genus Jndirana.
It should be confirmed whether the other
species which are allocated to Indirana have
the same sperm type.

The sperm head of Nyctibatrachus major
was very loosely coiled. The sperm shape
of Xenopus laevis (Bernardini et al., 1986,
1988; Yoshizaki, 1987) resembles that of N.
major. Some pelobatid frogs, Megophrys
montana (Asa and Phillips, 1988) and four
species of Vibrissaphora (Wu and Yang,
1981) have spermatozoa which are coiled
several times. According to Asa and Phil-
lipps (1988), chromatin condenses to form a
cylindrical coil within a spherical nucleus of
spermatids without participation of
microtubular manchette. The occurrence
of this sperm type in several unrelated anu-
ran families suggests a common cytological
basis for shaping this loosely coiled sperm
head. The coil structure of the sperm head
found in R. beddomii and Rhacophorus
species inhabiting east Asia may also have
the same basis. All types of coiled sperm
head reported so far are coiled sinistrally,
and this also suggests a common underlying

mechanism for producing coiled nuclear
condensation.

The sperm size of Rh. malabaricus is wi-
thin the range of east Asiatic Rhacophorus
species (Kuramoto, 1996). Contrary to the
compactly coiled sperm heads of these other
species, however, the sperm head of Rh.
malabaricus is straight. We could not ob-
serve even a trace of coiled structure at all.
It is necessary to examine the sperm shapes
of southeast Asiatic species to clarify sperm
variations within this genus. Sperm of
Polypedates maculatus is nearly the same as
those of east Asiatic Polypedates species in
both shape and size.

The crescent-like sperm heads of the ge-
nus Philautus are very similar to those of
Chirixalus species from east Asia (Kuramo-
to, 1996), suggesting a close relationship of
the two genera. The relationship from
spermatological data fits the phylogeny of
Channing (1989) fairly well, but not that of
Liem (1970). This shape of sperm head
may link straight and loosely coiled types.

Van der Horst et al. (1995) suggested a
correlation between sperm size and the
mode of fertilization in South African
frogs, that is, terrestrial fertilizers have long
spermatozoa and aquatic fertilizers short
ones. Kuramoto (1998), examining his
spermatological data, doubted this tenden-
cy. In the present materials, we observed
that a R. beddomii-like frog lays eggs in
seepages often exposed directly to the air
and the eggs were enveloped with tough
gelatinous coats. This mode of reproduc-
tion is not purely aquatic, so the longer
sperm of R. beddomii seems to fit the ten-
dency of van der Horst et al. (1995).
However, sperm of R. tagoi of Japan which
has a similar mode of reproduction and the
same kind of gelatinous envelope does not
differ from that of the other aquatic fertiliz-
ers (Kuramoto, 1998). The terrestrial
reproductive mode of Rhacophorus and
Polypedates seems to agree with the tenden-
cy, but that of Philautus seems to contradict
it. Although direct development in Philau-

KURAMOTO & JOSHY—FROG SPERM MORHOLOGY 69

tus was reported in species of Philippines
(Alcala and Brown, 1982) and Malaysia
(Yong et al., 1988) and not in Indian spe-
cies, Inger et al. (1984) collected most of
their Philautus specimens from places far
from any stream or pond. We also collect-
ed calling male specimens in forests or
bushes usually far from water. These sug-
gest a terrestrial mode of reproduction, but
sperm of Philautus are short. Rao (1937)
described larvae of some Philautus species
from streams of Kempholey, Hassan, Kar-
nataka. All have a flattened body, a long
muscular tail with a low tail fin, and large
mouth parts; all are characteristics of
stream-dwelling larvae. No information
about reproduction of Indian Philautus are
available, however. The presence of free-
living aquatic tadpoles does not necessarily
mean aquatic egg laying. The sperm size
does not seem to relate directly to the mode
of reproduction but seems to reflect
phylogenetic relationships fairly well.

ACKNOWLEDGMENTS

We thank Y. Nagayama and K.
Watanabe for SEM facilities, S. Rao, K. P.
Siju, P. Salins and S. Babu for aid in field
collecting, and Y. Shibata for literature.

LITERATURE CITED

AHL, E. 1931. Das Tierreich. Amphibia, Anura
III, Polypedatidae. Walter de Gruyter, Berlin.
477 p.

ALCALA, A.C. AND W.C. Brown. 1982.
Reproductive biology of some species of
Philautus (Rhacophoridae) and other Philip-
pine anurans. Phil. J. Biol. 11: 203-226.

AsA, C. S. AND D. M. Pgrrrrpps. 1988. Nuclear
shaping in spermatids of the Thai leaf frog
Megophrys montana. Anat. Rec. 220: 287-
290.

BERNARDINI, G., F. ANDRIETTI, M. CAMATINI
AND M.-P. Cosson. 1988. Xenopus sper-
matozoon: correlation between shape and
motility. Gamete Res. 20: 165-175.

BERNARDINI, G., R. STIPANI AND G. MELONE.
1986. The ultrastructure of Xenopus sper-

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Accepted: 22 April 2000

Current Herpetology 19(2): 71-79., Dec. 2000
© 2000 by The Herpetological Society of Japan

On the Monophyly of the Agamid Genus Gonocephalus Kaup,
1825 (Reptilia: Squamata): A Chromosomal Perspective

CHEONG-Hoonc DIONG!, May-Hon LOW!, EnE-CHoo TAN?, Hor-SEN
YONG3, Tsutomu HIKIDA‘, AND HipeEtTosu1 OTA5*

1 Division of Biology, School of Science, Nanyang Technological University, 469 Bukit
Timah Road, SINGAPORE 258756

2 Institute of Cell and Molecular Biology, National University of Singapore, SINGA-

PORE 117609
3 Institute of Biological Sciences, University of Malaya, 50603, Kuala Lumpur,

MALA YSIA

4 Department of Zoology, Graduate School of Science, Kyoto University, Kitashiraka-

wa, Sakyo-ku, Kyoto 606-8502, JAPAN
> Tropical Biosphere Research Center, University of the Ryukyus, Nishihara, Okinawa
903-0213, JAPAN

Abstract: We karyotyped five species of the agamid genus Gonocephalus, G.
chamaeleontinus, G. liogaster, G. bellii, G. grandis (from Peninsular Malay-
sia), and G. robinsonii. Of these, karyotypes of the first four species had
several chromosomal characteristics exclusively shared with the previously
reported karyotypes of G. miotympanum and G. grandis (from Borneo), such
as the diploid chromosome number (42) and the presence of 22 biarmed mac-
rochromosomes. This seems to support the monophyly of those four species
and G. miotympanum, probably along with some other species of the genus
not yet karyotyped. This hypothesis is premised on our finding of distinct
chromosomal characteristics that are indicative of highly derived states in the
agamid karyotypes. The karyotype of G. robinsonii, while remarkably differ-
ent from other congeneric karyotypes in exhibiting much smaller diploid (32)
and biarmed macrochromosome numbers (12), share these and other chro-
mosomal characteristics with several Australian species. It seems unlikely for
the karyotype of G. robinsonii to directly emerge from other congeneric
karyotypes or vice versa. We conclude that the inclusion of this species in
Gonocephalus would render the genus paraphyletic.

Key words: Reptilia; Agamidae; Gonocephalus belli; G. chamaeleontinus; G.
liogaster; G. grandis; G. robinsonii; Karyotype; Monophyly

___ < < 生ま INTRODUCTION
* Corresponding author. Tel: +81-98-895- ;

8937; Fax: +81-98-895-8966. The agamid genus Gonocephalus Kaup,
E-mail address: ota@sci.u-ryukyu.ac.jp (H. 1825, is a group of moderate-sized to large

Ota) lizards. Darlington (1957) argued that the

genus is zoogeographically exceptional, be-
cause it was then considered to occur on
both sides of Wallace’s Line, a zoogeo-
graphic border between the Oriental and
Australian faunas. Based on the micro-
chromosome numbers, Witten (1983) also
postulated that the Australian species as-
signed to Gonocephalus at that date
represent recent dispersals from Southeast
Asia across Wallace’s Line.

In his unpublished Ph.D. dissertation,
Moody (1980), on the basis of phylogenetic
analysis of morphological characters, as-
serted that the Gonocephalus species east of
Wallace’s line were derived from radiations
of the Australian stock, and that they are
phylogenetically distant from the Southeast
Asian species. He further argued that the
genus Hypsilurus Peters, 1867, once syn-
onymized to Gonocephalus by Boulenger
(1885), should be resurrected to accommo-
date species from the Australian Region.
Results of more recent immunogenetic
(Baverstock and Donnelan, 1990; King,
1990), karyological (Ota et al., 1992), elec-
tron-microscopic (Ananjeva and Mat-
veyeva-Dujsebayeva, 1996), and molecular
studies (Honda et al., 2000) favored Moo-
dy’s (1980) view. All recent authors, with
the exception of a few who have obviously
overlooked these works (e.g., Urban, 1999),
restrict the application of the generic name,
Gonocephalus, to the Southeast Asian spe-
cies (Welch et al., 1990; Manthey and
Grossmann, 1997).

In all those works addressing the
phylogeny of Gonocephalus (sensu lato),
however, the species assemblage on the
western side of Wallace’s Line Gr.e.,
Gonocephalus [sensu stricto]), though con-
stituting no less than 16 species (Welch et
al., 1990; Manthey and Grossmann, 1997),
was represented by very few species. For
example, Moody (1980) examined osteologi-
cal specimens for only five species. Fur-
thermore, Baverstock and Donnellan
(1990), Ota et al. (1992), Ananjeva and
Matveyeva-Dujsebayeva (1996), and Honda

Current Herpetol. 19(2) 2000

et al. (2000) examined only one, two, four,
and one species, respectively. Considering
that a thorough definition of Gonocephalus
(sensu stricto) depends only on a few exter-
nal characters (Moody, 1980; Manthey and
Grossmann, 1997), the monophyly of the
genus is obviously yet to be examined.

Ota et al. (1992) reported that G. grandis
and G. miotympanum, both from Borneo,
share characteristic chromosomal arrange-
ments that are obviously in highly derived
states. This suggests that the karyological
approach may be an effective way to exa-
mine the monophyly of the genus. There-
fore, in this study, we karyotyped four ad-
ditional species of Gonocephalus including
its type species, G. chamaeleontinus, as well
as G. grandis from the Peninsular Malaysia.

MATERIALS AND METHODS

Except for two male Gonocephalus
robinsonii, all lizards, collected from
Peninsular Malaysia and Pulau Tioman
(Table 1), were transported to the laborato-
ry where they were injected intraperitoneal-
ly with 0.1ml of colchicine solution
(2 mg/ml) per gram of body weight. Six-
teen to 18h after injection, they were
anesthesized with diethyl ether and were
dissected to remove femur bones. Bone
marrow were flushed out from the bones
with Hanks balanced buffer solution. For
each sample, the cell suspension was left to
stand for 10 min before it was centrifuged at
2000 rpm for Smin. Bone marrow cells
were then treated with hypotonic KCI solu-
tion (0.06 mole/1) at room temperature (26-
28°C) for 1 h, followed by fixation in a 1 : 3
glacial acetic acid : absolute methyl alcohol
mixture. Mitotic chromosome slides were
prepared by the splash technique, air-dried,
and were stained in 6% Gurr Giemsa (BDH)
solution. Mitotic cell slides for the remain-
ing two male G. robinsonii were prepared in
the field following Ota (1989a), and were
stained in 2% Giemsa solution.

DIONG ET AL.—PHYLOGENY OF AGAMID LIZARDS

TABLE 1.
mined in this study.

N
Species Males Females Total
G. bellii 2 3 5
G. chamaeleontinus 3 0 3
G. liogaster 0 3 3
G. grandis 3 3 6
G. robinsonii 3 0 3

た

Localities, sizes, and sexual compositions of samples of five Gonocephalus species exa-

Locality

Gombak Forest Reserve, Peninsular Malaysia
(03°09' N, 101?39 BE)

Pulau Tioman, near Peninsular Malaysia
(02?49 N, 104?09 BE)

Gombak Forest Reserve, Peninsular Malaysia
(03?09 N, 101?39 B)

Pulau Tioman, near Peninsular Malaysia
(02?49 N, 104?09 E)

Cameron Highlands Peninsular Malaysia
(04°28 N, 101°20’ EB)

Karyotypes were determined for each in-
dividual lizard on the basis of 8-20 well-
spread metaphase cells. Selected cell
spreads were photographed with a Nikon
Optiphot 2 Photomicrography camera us-
ing Kodak TMAX ASA 100 film. Individ-
ual chromosome pairs were arranged in
decreasing size. For the calculation of arm
ratio for each chromosome pair, the lengths
of chromosome arms were measured with a
CALCOM digitizer. Terminology for
chromosomal descriptions follows Green
and Sessions (1991), and the karyotype for-
mula follows Peccinini-Seale (1981).
Voucher specimens were deposited in the
Zoological Reference Collection, Depart-

ment of Biological Sciences, National
University of Singapore (ZRC) and Her-
petological Collection, Department of Zool-
ogy, Kyoto University (KUZ).

RESULTS

In all Gonocephalus species examined,
karyotypes consisted of chromosomes
forming large and smaller size-groups that
are referred to here as macrochromosomes
and microchromosomes, respectively (Table
2). Of these, macrochromosomes were all
bi-armed, whereas detailed morphology
remained undetermined for most micro-
chromosomes. No sex chromosome hetero-

TABLE 2. Karyotypes of species of the genus Gonocephalus. M=macrochromomes; m=

microchromosomes.
ae Chromosomal

Species 2n chromosomes formula Source

G. bellii 42 44 22M+20m this study

G. chamaeleontinus 42 44 22M+20m this study

G. liogaster 42 44 22M+20m this study

G. robinsonii 9 24 12 M 十 20 m this study

G. grandis (Pulau Tioman) 42 44 22M+20m this study

G. grandis (Borneo) 42 44 22M+20m Ota et al. (1992)
G. miotympanum 42 44 22M+20m Ota et al. (1992)

morphisms or secondary constrictions were
evident in any karyotypes.

Karyotypes of G. chamaeleontinus, G.
liogaster, G. bellii, and G. grandis consisted
of 2n=42 chromosomes, including 22
macrochromosomes (pairs 1-11) and
20 microchromosomes (pairs 12-21). The
macrochromosomes of G. chamaeleontinus
and G. liogaster were all metacentric (Fig.
1). From these, macrochromosomes of G.
bellii and G. grandis differed in including
submetacentric elements in pairs 1, 4 and
10, and pairs 2, 5, 7 and 9, respectively (Fig.
2). Thus, the arm numbers in macro-
chromosomes were 44 in all of the four
karyotypes. With respect to the micro-
chromosomes, the largest pair (i.e., pair 12)
of the G. bellii karyotype was distinctly en-
larged compared to the chromosome pair
immediately following, thus obscuring the
size-gap difference between the macro- and
microchromosomes. In contrast, chromo-
some pair 12 was almost as small as pair 13
in karyptypes of G. chamaeleontinus, G.
liogaster, and G. grandis, resulting in a
more prominent size-gap difference between
the two groups of chromosomes.

The karyotype of G. robinsonii differs
remarkably from those of the other four
Gonocephalus species in having substantial-
ly fewer (2n 三 32) chromosomes. Of the
diploid chromosomes, 12 (pairs 1-6) were
metacentric macrochromosomes, whereas
the remaining 20 (pairs 7-16) were micro-
chromosomes (Fig. 3). Therefore, the arm
number in macrochromosomes of this
karyotype equaled 24.

DISCUSSION

Karyotypes of G. chamaeleontinus, G.
bellii, G. liogaster, and G. grandis from
Peninsular Malaysia and Pulau Tioman
share similar chromosomal features with
those of G. miotympanum and G. grandis
from Borneo (Ota et al., 1992). In con-
trast, the karyotype of G. robinsonii differs
strikingly from other congeneric karyo-

Current Herpetol. 19(2) 2000

types.

In the family Agamidae, two karyo-
morphs are typical: (1) 2n=34 or 36 chro-
mosomes, including six pairs of metacentric
or submetacentric macrochromosomes and
11 or 12 pairs of microchromosomes; and
(2) 2n=46 or 48 chromosomes, all telocen-
tric chromosomes without a distinct size-
break (Bickham, 1984; King, 1981; Moody
and Hutterer, 1978; Olmo, 1986; Ota and
Hikida, 1989; Solleder and Schmid, 1988;
Witten, 1983). One of these karyomorphs
is considered to be derived from the other
through a series of Robertsonian rearrange-
ments of macrochromosomes, sometimes
accompanied with addition or deletion of
one microchromosome pair (Bickham,
1984; King, 1981). Judging from the fact
that both of the two karyomorphs some-
times occur in a single genus or closely
related genera (Gorman and Shochat, 1972;
Ota, 1988, 1989b) and that there are so few
karyotypes representing intermediate states
between the two extremes, such chro-

mosomal rearrangements may proceed
rapidly when they are once triggered (King,
1981).

It is obvious that the karyotypes of G.
chamaeleontinus, G. bellii, G. liogaster, G.
grandis, and G. miotympanum could not be
derived from either of the two typical
agamid karyomorphs merely through
Robertosonian rearrangements of macro-
chromosomes and slight numerical changes
of microchromosomes, because _ these
karyotypes include a much larger number of
biarmed macrochromosomes (22) than
those of any other agamid karyotypes
hitherto reported, and at the same time, ex-
hibit a greater diploid number (42) than the
agamid karyomorph (1). Moreover, their
fundamental number (NF, >64) is con-
siderably greater than those with karyo-
morph (2) (NF=46 or 48). We thus con-
sider karyotypes of the five Gonocephalus
species to represent a highly derived state,
and that the chromosomal features exclu-
sively shared among these species support

DIONG ET AL.—PHYLOGENY OF AGAMID LIZARDS 5

A
a 76
] 2 3 4 5 6 7
8 9 10 f ルレ 6b 1 5
ee る ee © es
6 21
トーーーーーーーー ゴ 1

ーー | mwl に ーー ーー

Fic. 1. Karyotypes of (A) Gonocephalus chamaeleontinus, and (B) G. liogaster. Bars equal 5 4m.

76 Current Herpetol. 19(2) 2000

3 EE

] 2 3 4 り

まま @@ 48 66
; o 10 1 2 8 4 oa

i 8 9 10 i レ 8 4 Ss
*? クッ 4 ク on m4
ee =〈。 の
トーーーーーーーー ゴ

Fic. 2. Karyotypes of (A) Gonocephalus bellii, and (B) G. grandis. Bars equal 5 ym.

DIONG ET AL.—PHYLOGENY OF AGAMID LIZARDS a7

S®e@ “24 @6 @0@ © 06 ©60 *# «* «ae

Fic. 3.

their monophyly, presumably along with
some other species of the genus not yet stu-
died karyologically.

The karyotype of G. robinsonii is similar
to karyomorph (1), and _ considering
remarkable differences in the diploid num-
ber of chromosomes or the arm number of
macrochromosomes between this and other
congeneric karyotypes (Table 2), it is un-
likely that the karyotype of G. robinsonii
directly arose from other congeneric karyo-
types or vice versa. Thus, we conclude that
the inclusion of this species in Gonocepha-
lus would render the genus paraphyletic.

The number of microchromosomes (20)
in the G. robinsonii karyotype is smaller
than that in the typical agamid karyomorph
(1)(22 or 24: Bickham, 1984; King, 1981).
Such a chromosomal arrangement (1.e.,
12M+20m) is exclusively shared with
several agamid species that are supposedly
derived from the Australian endemic radia-
tion (Witten, 1983). Itis thus probable that

Karyotype of Gonocephalus robinsonii. Bar equals 5 um.

G. robinsonii actually represents dispersals
from the Australian Region into Southeast
Asia like Physignathus cocincinus (see Hon-
da et al., 2000). However, it is also proba-
ble that the karyotype of G. robinsonii is
derived from karyomorph (1), exhibited by
a number of other agamid species including
several from Southeast Asia (see Ota and
Hikida [1989]), through deletions of one or
two microchromosome pairs. More com-
prehensive analyses using biochemical and
molecular approaches are needed to deter-
mine the relationship of this enigmatic spe-
cies with certainty.

ACKNOWLEDGMENTS

We thank the Institute of Biological
Sciences, University of Malaya, Kuala
Lumpur, for provision of facilities for ex-
periments. We are also much indebted to
J. A. Schulze for commenting on an early
version of the manuscript. H. Ota and T.

Current Herpetol. 19(2) 2000

Hikida are grateful to M. Matsui and K.
Araya for helping with the fieldwork in the
Cameron Highlands, Peninsular Malaysia.
This study was partially supported by an
academic research grant from the Nanyang
Technological University of Singapore
(No. RP21/97, to CHD) and a Grant-in-Aid
from the Japan Ministry of Education,
Science, Sports and Culture (No. 0404068
to M. Matsui).

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© 2000 by The Herpetological Society of Japan

Validity of Back-calculation Methods of Body Size from
Phalangeal Bones: An Assesment Using Data for
Rana japonica

JUNSUKE MARUNOUCHI'"*, Tamotsu KUSANO? ANp Hiroaki UEDA!**

1 Laboratory for Amphibian Biology, Graduate School of Science, Hiroshima Univer-

sity, Kagamiyama 1-3-1, Higashi-Hiroshima 739-8526, JAPAN

2 Department of Biology, Faculty of Science, Tokyo Metropolitan University,

Minamiohsawa, Hachioji, Tokyo 192-0397, JAPAN

Abstract: Lines of arrested growth (LAGs) appearing in bone sections are
useful for age estimation. They also indicate the past growth process in am-
phibians in temperate zones. Several back-calculation formulae (BCFs) use
LAGs to estimate an individual’s body size at an earlier time based on the
current body size. In order to evaluate the validity of these BCFs, we con-
ducted a mark-recapture and skeletochronological study of female Rana
japonica in Higashi-Hiroshima, Japan, from 1995 to 1999. The body sizes of
31 recaptured frogs were back-calculated using eight different BCFs and were
compared with the frogs’ actual body sizes as measured at the previous cap-
ture. The most accurate estimation was made by the simplest BCF (Dahl-Lea
method) without any regressions between body size and bone diameter; that is,
L,=L,.(D,/D,) (L: snout-vent length, D: bone diameter, c: at the time of cap-
ture [recapture], i: at the i-th winter).

Key words:
japonica

INTRODUCTION

In the study of life history and population
dynamics, it is important to know the
processes of growth and sexual maturation.
However, it is difficult to follow an individ-
ual organism’s growth throughout its life by
the mark-recapture method, especially in
the case of long-lived species such as some
amphibians.

* Corresponding author.

E-mail address: marujun@hiroshima-u.ac.jp
(J. Matunouchi)

* HIROAKI UEDA was deceased in 1997.

Skeletochronology; Body size; Back-calculation formula; Rana

Skeletochronology has proven to be an
effective method of age estimation for am-
phibians in the temperate zone (see Halliday
and Verrel, 1988; Castanet and Smirina,
1990). Lines of arrested growth (LAGs) are
formed annually in the bone at periods of
growth retardation such as hibernation.
Thus, LAGs provide valuable information
not only on the organism’s age (the number
of winter seasons a given individual has ex-
perienced), but also on the process of bone
growth with age (Hemelaar, 1988; Fretey
and Le Garff, 1992; Neveu, 1992) and sexual
maturation (Francillon-Vieillot et al., 1990;
Augert, 1992; Augert and Joly, 1993;

Current Herpetol. 19(2) 2000

Caetano and Castanet, 1993; Kusano et al.,
1991, 1995a, b).

Since bone growth is considered to reflect
growth in body size, body size at an earlier
time can be back-calculated from the size of
the zone encompassed by a LAG and the
current body size (Smirina, 1983; Ryser,
1988; Augert, 1992; Neveu, 1992; Augert
and Joly, 1993; Caetano and Leclair, 1996;
Leclair and Laurin, 1996). Several different
back-calculation formulae (BCFs) have
been proposed for the estimation of body
size at a past time based on a certain linear
relationship between bone size and body
size (see Francis, 1990; Ricker, 1992).
Although Ryser (1988) commented briefly
on the validity of two BCFs for frogs, the
validity of these BCFs has not yet been
evaluated sufficiently for amphibian species.

We conducted a mark-recapture and
skeletochronological study on a population
of the Japanese brown frog, Rana japonica.
Applying different BCFs to recaptured
frogs, their body sizes at the previous cap-
ture were estimated, and the back-calculat-
ed sizes were compared with the actual sizes
in order to assess the accuracy of the BCFs.
Here, we report the results of the evaluation
and discuss the validity of these BCFs as
applied to frogs.

MATERIALS AND METHODS

The Japanese brown frog, Rana japoni-
ca, is commonly distributed in Honshu,
Shikoku, and Kyushu, and breeds from
January to April in still waters such as rice
fields, marshes, and small pools (Maeda and
Matsui, 1989). We studied a population of
R. japonica at “Hiroshima University Eco-
logical Garden” (34°24' N, 132°43’E, alti-
tude 220m) within the campus of Hiroshi-
ma University in Higashi-Hiroshima,
Hiroshima Prefecture, Japan.

The mark-recapture study was conducted
chiefly during the breeding seasons (Janu-
ary-March) from 1995 to 1999. Since 1996,
the survey has also been conducted in a

non-breeding period as well (September-
November). Frogs were captured by hand
or using a dip net. They were then meas-
ured for snout-vent length (SVL) to the
nearest 1 mm with a slide caliper, and sexed
on the basis of secondary sexual charac-
teristics such as the development of thum-
bpads and ovaries. The frogs were individ-
ually marked by toe-clipping, and were
released at the capture sites. Group mark-
ing was applied to juveniles newly
metamorphosed in 1997-1998, since they
were too small to be individually marked.
When a frog was captured, the fourth toe of
the left or right hind leg was clipped off for
a Skeletochronological study. Each toe was
decalcified in 6% nitric acid for 40 min and
washed in running tap water for 24h. Af-
ter being embedded in paraffin, the second
phalanx of each toe was sectioned (15 um
thick) and stained with Lili-Meyer’s
hematoxylin. We selected the best cross-
section for each individual, and measured
the major and minor axes of the outer mar-
gin of the bone and LAGs using an ocular
micrometer attached to a light microscope.
Originally we used two kinds of bone di-
ameter: the length of the major axis, and the
geometric mean of the major and minor
axes. Since the results of the analyses using
each kind of diameter were quite similar, we
report here the results of the analysis using
the major axis only.

We assumed that the SVL and phalangeal
size of individuals captured in autumn
would be nearly equal to those in hiberna-
tion, because the mark-recapture study
showed that most of the frogs did not show
any apparent growth until the next spring.
Therefore, we pooled data for yearlings
captured in late autumn with those for one-
year-olds captured in spring for the analy-
sis. In an earlier study, Francis (1990) rev-
iewed the literature on the calculation of
fish lengths at successive ages from growth
marks on hard parts of the body, such as
scales and otoliths, and he presented several
typical BCFs that had been applied in past

MARUNOUCHI ET AL.—FROG SIZE ESTIMATION 83

researches. Although Francis (1990)
recommended an ordinary regression (OR),
Ricker (1992) proposed that the geometric
mean regression (GMR, also called the
reduced major axis) between bone diameter
and body size be used as a regression equa-
tion in BCFs. In the present study, both
types of regressions were used in BCFs.
Based on the earlier researches, we applied
the following eight BCFs to R. japonica (see
Francis, 1990; Ricker, 1992).
(1) Dahl-Lea method: L;=L,D;,/D,
(2) Regression method :
(2-1) L.=p+qD; (OR)
(2-2) L,=u+vD,; (GMR)

(3) Fraser-Lee method :

(3-1-1) L;=(L,+a/b)D;/D,-a/b (SPH
method using OR; Fran-
cis, 1990)

(3-1-2) Li=(L.-p)Dj/D, + p (OR)

(3-2) L;=(L,-u)D;/D, + u (GMR)

(4) Whitney-Carlander method :

(4-1) Li=L.(p+qDj)/(p+qD.) (BPH
method using OR; Francis,
1990)

(4-2) L,=L,(u+vD;)/(u+ vD,) (GMR)

L: SVL in mm; D: bone diameter in ym;
c: at capture (recapture in the present
study); i: at i-th winter; a, b: D-intercept
and slope, respectively, from the ordinary
regression equation, D=a+DbL,; p, q: L-in-
tercept and slope, respectively, from the or-
dinary regression equation, L=p+qD; u,
v: L-intercept and slope, respectively, from
the geometric mean regression (GMR) equa-
ton, L—u-_vb.

We used three regressions between SVL
and bone diameter in this analysis, as men-
tioned above, and these regression equa-
tions were calculated from the sample of 74
females in the breeding seasons of 1995 and
1996 and eight juveniles in the autumn of
1996 (see Fig. 1). For recaptured frogs,
their body sizes at first capture were esti-
mated using different BCFs, and the back-
calculated sizes were compared with the ac-
tual sizes. For each BCF, the difference be-
tween the back-calculated sizes and the ac-

SVL (mm)

0 100 200 300 400 500
Bone diameter ( um)

Fic. 1. The relationship between snout-vent
lengths (SVL or L) and bone diameter (D) of
phalanges. Open circles and squares indicate
adults and juveniles, respectively. Three types
of regressions are shown: the dotted line is an
ordinary regression (OR) of D on L, D=
— 32.77+7.72L (r=0.89, p く 0.001): the narrow-
er solid line is an OR of L on D, L=
13.43+0.10D; the broader line is a geometric
mean regression (GMR), L=9.11+0.12D.

tual sizes was evaluated using the paired t-
test at a significance level of 5%. Since the
main purpose of this study was to detect the
estimate error in order to evaluate the ac-
curacy of BCFs, we placed a high value on
the power of the test by minimizing the type
II error rather than the type I error (Sokal
and Rohlf, 1995; Jaeger and Halliday,
1998). Therefore, we did not adjust p-
values, even when multiple tests were per-
formed.

RESULTS

During the study period, a total of 31 fe-
males were recaptured between seasons, but
very few males were recaptured. There-
fore, we report only on the results for the
females. The 31 recaptured females had
first been captured in the period between
February, 1996, and February, 1999, and

SVL at recapture (mm)

30 40 50 60 70
SVL at first capture (mm)

Fic. 2. The relationship between SVLs at
recapture (R) and at first capture (F). The solid
line indicates the regression line, R=50.53
+0.24F (r=0.53, p=0.002) and the dotted line
is an isoline of F=R.

し

SN Ns

7 ve 1
ie

ーー

Fic. 3.

Current Herpetol. 19(2) 2000

they were recaptured between February,
1997, and September, 1999. No individuals
experienced more than one active season be-
tween first capture and recapture. Their
SVLs ranged from 30 to 58 mm (mean+SD,
40.1 土 7.7) at first capture, and from 55 to
67 mm (60.2+3.5) at recapture (Fig. 2).
Twenty-eight of the 31 frogs were two
years old, and the other three frogs were
three years old at recapture. At first cap-
ture, 18 frogs were sexually mature, whereas
13 were juveniles. In five of the 18 adults
and in all 13 juveniles, phalangeal bones
were obtained only at recapture, and
skeletochronological observation of cross-
sections showed that they all were two years
old at the time of recapture. Inthe other 13
adults, phalanges were collected at both first
and second captures. These adults had one
more LAG at recapture than they had at

Cross-sections of phalanges at first capture (left) and recapture (right). The black bar in

each photograph indicates a scale of 0.1 mm. (A): A one-year-old female of 40 mm SVL on 11 March

1998.

(B): The same individual as (A) measuring 58mm SVL on 10 February 1999.

(C): A two-

year-old female of 55 mm SVL on 8 February 1996. (D) The same individual as (C) measuring 59 mm

SVL on 16 February 1997.

MARUNOUCHI ET AL.—FROG SIZE ESTIMATION 85

first capture, which corresponded to a single
hibernation (Fig. 3).

The mark-recapture data for these fe-
males were analyzed in order to assess the
accuracy of the BCFs. Table 1 shows the
results of the comparisons between the
back-calculated sizes and the actual sizes.
The mean SVLs back-calculated by respec-
tive BCFs ranged from 40.7 to 45.2 mm,
and were more or less larger than the actual
value of 40.1 mm (Table 1). The smallest
mean of absolute error in the BCFs was
2.3mm in BCF (1), and the largest value
was 5.2mm in BCF (4-1). Significant
differences were detected between back-cal-
culated and actual SVLs in all BCFs except
BCF (1) (paired t-test, p<0.05). The
results showed that among the eight BCFs,
BCF (1) gave the most accurate estimate,
although it was the simplest equation and
did not employ any regressions.

The relationships between the estimates

and the actual SVLs are shown in Fig. 4.
Only according to BCF (1), which was the
best BCF among those examined (see
above), the regression between the estimates
(F.) and actual SVLs (F) was not sig-
nificantly different from the isoline of y=x.
The regressions for other BCFs exibited sig-
nificant deviations from the isoline. In
BCFs (3) and (4), the estimate error became
larger when the SVL was smaller, while the
estimates were relatively close to actual
values at large SVLs. On the other hand,
the estimate errors increased at both ex-
treme values of SVL, and were relatively
small at medium SVLs in BCF (2) (Fig. 4).
In all BCFs except for BCF (1), there were
significant negative correlations between es-
timate error (E) and actual SVL at first cap-
ture (F), rrE (p<0.05, Table 2). We ana-
lyzed other correlations among the variables
E, F, actual SVL at recapture (R), and
growth (G) to determine the estimate biases

TABLE 1. Results of back-calculation of snout-vent lengths (SVLs) at the previous capture based
on eight back-calculation formulae (BCFs). The difference between estimated SVLs and actual SVLs
are tested by the paired t-test. Asterisks denote statistically significant differences (p<0.05).

SVLs (mm) Ratio
x+SD x 士 SD
BCF (range) (range)
Actual 40.1+7.7
value (30-58)
(1) 40.7+7.4 1.02+0.08
2(29-57) (0.89-0.22)
2-1) 42.7 土 5.4 1.08+0.10
(36-57) (0.88-1.30)
+ +
(2-2) 42.1+6.1 1.06+0.09
(34-58) (0.86-1.27)
+ +
(3-1-1) 42.0+6.9 1.06+0.08
(31-57) (0.91-1.25)
45.0+5.9 1.14+0.10
3-1-2
( ) (35-58) (0.98-1.37)
+ +
(3-2) 43.5+6.4 1.10+0.09
(33-57) (0.98-1.31)
(4-1) 45.2+6.1 1.14+0.11
(35-58) (0.98-1.37)
a +
(4-2) 43.6+6.5 1.10+0.09
(33-58) (0.97-1.31)

Error (mm) Absolute error (mm) Paired
x 士 SD x 土 SD t-test
(range) (range) p-value

ais pelea lls
ne 0213
+ +
Zh ange Cater
ais Gye A,
eC Sa
コピ POD
ae の Se 0 0.001*
ote aster
ee ee
+ +2.
Sieg evs gigas od
+ +
See PMS <
+ +
BPYP ay nlaa<0001"

Estimates (mm)

30 ’ 40 50 5. 30 40 50 60
Actual SVL (mm)

Fic. 4. Therelationship between back-calcu-
lated (F。) and actual SVLs (F). Open circles and
squares indicate adults and juveniles at first cap-
ture, respectively. Solid lines indicate regression
lines of F。 against F and dotted lines are isolines
of F.=F. (A): BCF(1) F.=4.98 + 0.89F (r=0.93,
p く 0.001): (B): BCF(2-1) F.=17.17+0.64F
(r=0.90, p く 0.001): (C): BCF(2-2) F.=13.55+
0.71F (r=0.90, p<0.001); (D): BCF(3-2-1)
F.=8.66+0.83F (r=0.93, p く 0.001): (E): BCF
(3-2-2), F.=16.50+0.71F (r=0.93, p<0.001);
(F): BCF(3-2), F.=12.57+0.77F (r=0.93, p<
0.001); (G): BCF(4-1), F.=16.36+0.72F (r=0.91,
p<0.001); (H): BCF(4-2) F,=12.17+0.79F
(r=0.93, p<0.001). See text for abbreviations
for back-calculation formulae.

of the respective BCFs (Table 2). In BCF
(2-1), rsE was significantly negative, indicat-
ing Lee’s phenomenon in which the larger

Current Herpetol. 19(2) 2000

individuals at the last capture were estimat-
ed to be smaller at any past age (Ricker,
1992). In all of the BCFs, a positive ras
(p<0.05) showed that the estimates of past
SVLs of fast-growing individuals were larg-
er than actual. However there was a sig-
nificantly positive correlation between R
and F (rrs 三 0.33, p=0.002, Fig. 2), and a
negative correlation between G and F (rra
= — 0.89, p<0.001). Therefore, we also
analyzed partial correlations among these
variables.

In all BCFs, the partial correlations con-
trolled for R (rr .s) were significantly nega-
tive (p<0.05). In contrast to rpg, the par-
tial correlation rs .r indicated the inverse of
Lee’s phenomenon in BCFs (1), (3), and (4).
These BCFs also showed the tendency to
overestimate past SVLs of fast-growing in-
dividuals (significantly positive ra .r). The
Dahl-Lea, Fraser-Lee, and Whitney-Carlan-
der methods were thus affected by the size
of the frog at last capture and by growth.

We collected as many juveniles as possi-
ble and measured their SVLs in autumn of
1996 and 1997, in order to determine the
minimum SVL at sexual maturity. Their
SVLs ranged from 24 to 40mm, with the
mean being 31.0 (SD=3.3, N=99). Since
the upper limit of the 95% range based on
t-distribution was 37.4mm when back-cal-
culated SVLs were 37 mm or less, the frogs
were estimated to have been juveniles at the
time of capture. The number of individ-
uals estimated to have been juveniles based
on back-calculated SVLs varied greatly
from 2-10 depending on the BCF used
(Table 3). BCF (1) gave a value of 10, and
its rate of correct judgement was 0.83 as a
whole, the best performance among the
BCFs examined. The other BCFs gave very
poor judgement of juveniles that were less
than half of the actual number in most
cases. BCFs (3-1-2) and (4-1) gave the
lowest estimate of only two. The BCFs
other than BCF (1) are considered to be in-
effective in providing this estimate.

MARUNOUCHI ET AL.—FROG SIZE ESTIMATION 87

TABLE 2.
(p<0.05).

Correlations between estimate errors and SVLs. Asterisks show significant correlations
Symbols of variables are: F, actual SVL at first capture; R, actual SVL at recapture; G,

growth, i.e., difference of SVL between captures; E: estimate error, i.e., difference between estimates

and actual SVL at first capture.

BCF TEB IFE.R
(1) — 0.30 =—0-45*
(2-1) —0.77* ー0.72*
(2-2) ー0.6$* —0.60*
(3-1-1) =0.46* —0.60*
(3-1-2) —0.71* —0.80*
(3-2) —0.59* —0.70*
(4-1) — 0.66* —0.79*
(4-2) —0.55* —0.67*
TABLE 3.

N of individuals
judged correctly
BCF as juveniles
Actual N 13
(1) 10
(2-1)
(2-2)
(3-1-1)
(3-1-2)
(3-2)
(4-1)
(4-2)

AN nan OD 選

DISCUSSION

In almost all of the phalangeal cross-sec-
tions of R. japonica, stained LAGs were
distinct, being easily counted and measured
(Fig. 3). The mark-recapture procedure
showed that skeletochronology was an
effective age-determination method in this
species in addition to some other amphibian
species that had been studied previously,
such as Bufo bufo (Hemalaar and Van Gel-
der, 1980; Fretey and Le Garff, 1992), R.
temporaria (Gibbons and MaCarthy, 1983),
and B. calamita (Tejedo et al., 1997).

Francis (1990), based on the survey of a
number of relevant papers on fish popula-

TRE TRE.F IGE IGE.F
0.14 Onis 0.43* Ome
053° 0.04 0:70" 0.04
ー0.30 0.06 0.60* 0.06
0.09 0.44* 0.59* 0.44*
ー0.06 O25 0.80* 0:52"
0.00 0.46* 0.70* 0.46*
0.02 Oa: 0.78* 0577
0.02 0.44* 0.66* 0.44*

Validity of determination of sexual maturity based on back-calculated SVLs.

N of individuals

judged correctly Rate of correct

as adults judgements

18 bes

16 0.83 (26/31)
18 0.71 (22/31)
17 0.74 (23/31)
16 0.77 (24/31)
18 0.65 (20/31)
17 0.71 (22/31)
18 0.65 (20/31)
17 0.71 (22/31)

tions, demonstrated that the Fraser-Lee
method (BCF (3)) was the most popular,
followed by the regression (BCF (2)) and
Dahl-Lea (BCF (1)) methods in that order.
Ricker (1992) evaluated the validity of BCFs
on theoretical grounds, and argued that the
regression between scale (bone) diameter
and body length for BCFs must be GMR
rather than OR, and that the Fraser-Lee
method (BCF (3-2)) is preferable.

Similar studies of amphibian species in-
clude those of Kusano et al. (1991), who
applied the Dahl-Lea method (BCF (1)) to
B. marinus, and Augert and Jolly (1993),
who used the regression method (BCE (2-1))
for R. temporaria. The Fraser-Lee method

has also been used for back-calculation in
amphibian species. For example, BCF (3-
1-2) has been applied to B. bufo (Smirina,
1983), R. septentrionaris (Leclair and Lau-
rin, 1996) and Notophthalmus viridescens
(Caetano and Leclair, 1996). Neveu (1992)
reported that the mean body size of a cer-
tain age group, which was estimated by the
Whitney-Carlander method (BCF (4-1)),
did not differ from that derived from the
regression method (BCF (2-1)) for R. es-
culenta. In R. temporaria, Ryser (1988)
used not only an ordinary regression line
between SVL and phalangeal diameter, but
also a specially devised linear equation. He
assesed the accuracy of BCF (2-1) and BCF
(4-1) by comparing values estimated by
these methods and actual values from
mark-recaptures. His results suggested that
BCF (4-1) was preferable, since the mean of
absolute error was 2.13 mm as compared to
3.13 mm for BCF (2-1).

In the present study, the accuracy was
compared between the BCFs incorporating
OR and GMR as regression procedures:
e.g., between (2-1) and (2-2), (3-1-2) and
(3-2), and (4-1) and (4-2). The results
showed that BCFs employing GMR give
more accurate estimates than those using
OR (Table 1). This supports Ricker’s
(1992) claim that GMR is preferable to OR
for BCFs. The Fraser-Lee method (BCF
(3)), exclusive of the SPH option (BCF (3-
1-1)), and the Whitney-Carlander method
(BCF (4)) made larger errors than BCF (2),
regardless of regression methods. BCF(1)
is just a special form of BCFs (3) and (4): it
is identical to BCFs (3) and (4) if y-inter-
cepts (a, p, and u) are fixed at zero. Stan-
dard regression theory shows that -a/b is al-
ways less than p (Francis, 1990). There-
fore, the estimates made by BCF (3-1-1)
were nearer to those of BCF (1) than of
BCF (3-1-2), but BCF (3-1-1) was still less
accurate than BCF (1). Therefore, at least
for R. japonica, the relationship between
SVL and phalangeal diameter seems to be
inappropriate for the “scale proportional

Current Herpetol. 19(2) 2000

hypophysis (SPH)” and the “body propor-
tional hypothesis (BPH)” as had been sug-
gested by Francis (1990) for fishes.

In the present study, the best BCF among
the eight different BCFs examined proved to
be the simplest formula using the Dahl-Lee
method (BCF (1)) (Table 1). All of the
other BCFs used the regression equations
between bone diameter and SVL: only BCF
(1) did not require such _ regressions.
Despite such simplicity, BCF (1) gave the
most accurate estimates. The mean of ab-
solute error for BCF (1) was 2.3 mm, only
about 6% of the actual SVL (Table 1). The
formula’s simplicity is also advantageous
for its application to population studies in
the field. Although a large sample of target
animals with a wide range of body sizes or
ages is needed to calculate reliable regres-
sions, BCF (1) does not require such a large
sample. This BCF can be applied even to a
small sample and still produce estimates
with high accuracy.

ACKNOWLEDGEMENTS

We thank Gentaro Toyohara, Tsuneo
Shioji, and Hirohide Ueki of the Depart-
ment of Biological Science, Faculty of
Science, Hiroshima University, for allowing
us to study in the Ecological Garden. We
also thank Masayuki Sumida and colleagues
from our laboratory as well as members of
the Laboratory of Aquatic Ecology, Faculty
of Applied Biological Science, Hiroshima
University, for their valuable advice.

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Accepted: 22 August 2000

OTT AMITEe Site

Current Herpetology 19(2): 91-96., Dec. 2000
© 2000 by The Herpetological Society of Japan

Relationship Between Brightness or Size and the Presence of
Barnacles on the Carapace of the Hawksbill Turtles

(Eretmochelys imbricata)

Mari KOBAYASHI

Laboratory of Wildlife Biology, Department of Environmental Veterinary Sciences,
Graduate School of Veterinary Medicine, Hokkaido University N18 W9, Kita-ku,

Sapporo 060-0818, JAPAN

Abstract: In this study, by converting the colors of the first coastal scutes
(C1) in the hawksbill turtle, Eretmochelys imbricata, into numerical values by
the shade (256 phases), it became possible to fully analyze the brightness of the
carapace. Two thousand seven hundred fifty-six C1s of hawksbill turtles that
had been captured in the Cuban sea from 1993 through 1994 were used. The
relationship between the brightness and the width of the first costal (C1W) was
examined. The results showed no correlation between brightness and C1W.
However, C1 with barnacles tends to have greater C1W (27.1 土 2.8 cm), while
no barnacles were found on C1 with a width of 20.3 cm or less. C1 with bar-
nacles, compared to C1 without, was low in brightness (somewhat dark) in

terms of statistical significance.

Key words:

INTRODUCTION

It has been reported that changes in pat-
tern as the carapace grows help to roughly
classify the sex or life stages of several turtle
species (Barbour and Carr, 1940; Moll,
1980). Although it is not clear whether this
fact is linked to the age of the turtle, it is
highly likely that it is related to the sex and
size (McCoy, 1968; Balazs, 1986). There
have been no studies on this relationship in
sea turtles, because it is difficult to track
turtles in the sea to study their carapace
patterns. While the carapace is made of
hard tissues and very likely to remain after
death from a cause like stranding. So it is
interesting to obtain new information on its
color or pattern from the carapace.

Hawksbill turtle; Barnacle; Brightness; Scute; Size

In this study, the Cl brightness of
hawksbill turtles, Eretmochelys imbricata,
living in the Cuban sea was converted into
numerical values by 256 phases of shade,
and then its relationship to the width of the
first coastal (C1W) was examined. Next,
between the turtle groups with barnacles or
not, the relationship between C1W and the
brightness was studied. There has been no
information on either the size or the bright-
ness of the hawksbill carapace. Not only
that, there have been no reports on the rela-
tion of barnacles attached to the carapace
and the brightness of the carapace.

MATERIALS AND METHODS

I used the right first costal (Cl) of

Current Herpetol. 19(2) 2000

hawksbills (N=2,756) that were captured
using fishing top nets of 46-53 cm mesh, 50-
60 fathoms long, 12-15 meshes deep, from
1993 to 1994. The sex, carapace length,
and other information from each turtles
were unknown. The reason for selecting
C1 is that C1 is very distinct from all other
scutes in shape, and therefore it is easy to
distinguish. Furthermore, by using only
the left side of Cl, a redundant use of tur-
tles can be avoided.

I first studied the relationship between the
brightness of C1 and C1W was examined.
Then, for individual groups, whether with
or without barnacles (Chelonibia), I studied
the relationships between C1W and the
brightness. Regarding barnacles on log-
gerheads (Caretta caretta), there are reports
that they attached themselves to the dorsal
carapace more than the ventral carapace
(Gramentz, 1988) and attachment on the
ventral carapace could be seen on the an-
terior, posterior, and middle vertebral
scutes (Matsuura and Nakamura, 1993).
But since hawksbills have the habit of dig-
ging under a coral reef and it is considered
that barnacles on the middle vertebral
scutes can be easily scraped off, I chose Cl
to examine whether there were attached
barnacles or not. The two groups were
defined as turtles in which traces of attached
barnacles could be observed, and turtles
with barnacles attached to them.

Brightness of Cl

Colors are generally classified by the three
basic characteristics of hue, intensity, and
brightness. Hue is expressed by the
wavelength of light. Intensity indicates the
degree of saturation or dullness of the color
and is expressed by the amount of gray
proportionate to hue. Brightness indicates
the degree of shade of the color ranging
from 0% (black) to 100% (white). In this
study, I divided the degree of brightness
into 256 computer-classifiable phases, con-
verting Cl into numerical values of shade.

With the newly formed scute side facing

upwards, photographs of Cl were shot ina
display case and a spot light on two separate
days under same shooting and film-develop-
ing conditions. Using a scanner, the pho-
tographs were converted into image data
and the brightness of Cl was converted into
numerical values using the degree of shade
(256 phases). In other words, the numeri-
cal value of shade was high on the amber-
colored part while the black speckles were
low in the numerical value of shade. I used
Image8.3 by Erdas (Inc.) for analyzing the
shade of the colors. For all the turtles, I
determined the mean brightness (“bright-
ness” hereafter) of the entire C1 area.

C1W
C1W was individually measured using a
tape measure (+0.1 cm).

Statistical treatment

The correlation between the Cl bright-
ness and the C1W was examined using
regression lines. The relationships between
the classification depending on the presence
of barnacles, C1W, and the brightness were
clarified by a one-way layout ANOVA
without assuming homogeneity.

RESULTS

Relationship between brightness and ClW

No correlation was observed between the
brightness and C1W (Fig. 1). The regres-
sion slope was Y=0.3845X + 92.133 and r?
was 0.0026.

Relationships between the presence of at-
tached barnacles and C1W or the brightness

It became clear that there is no correla-
tion between the brightness and C1W (Fig.
1). Thus, I compared C1W and the bright-
ness between the groups with (N=669) and
without barnacles (N=2087), through a
one-way layout ANOVA (Tamhane)
without assuming homogeneity. The
results showed that there was a significant
difference in C1W depending on the

KOBAY ASHI—CARAPACE OF HAWKSBILL TURTLE 93

の
の
®
=
tS
9
oo
15.0 20.0 25.0 30.0 35.0 40.0
C1W (cm)
Fic. 1. Scatter plots of C1W (in cm) and brightness (in 256 phases, N=2,756).

presence of barnacles (F=135.1, p く 0.01).
The mean C1W with barnacles was
27.142.8cm while without barnacles the
mean was 25.5+3.2cm. The frequency
distribution of C1W with or without barna-
cles is clearly different (Fig. 2). C1W of
20.3 cm or smaller showed no barnacles. In
the same manner, the relationship between

180
160
140
120
100

Number of turtles

17 19 21 23 25

barnacles and the brightness indicated that,
compared to others, Cl with barnacles is
significantly low in brightness (somewhat
dark) (F=22.2, p く 0.01). The mean
brightness of Cl with barnacles was
98.3+25.5 and the rest was 103.3 土 23.6.

Without barnacles
Mi «6UWWith barnacles

27 29 31 33 35 37 39
C1W (cm)

Fic. 2. C1W distibution of turtles with (dark rectangles) and without (gray rectangles) barnacles.

DISCUSSION

Melanin that has been accumulated on
melanocytes is converted into granule cell
melanosomes, and then shifts to the
neighboring cell keratinocytes that produce
hawksbill scutes, gradually forming a cara-
pace through keratinization. When this
happens, the black and amber colors shift
to keratinocytes simultaneously. Since in-
tracellular melanin consists of eumelanin of
black pigment (Seiji, 1970; Oikawa, 1976)
and pheomelanin of yellow-to-red pigment
(Prota, 1980), the former is considered to
make black speckles, and the latter amber
areas, thereby giving different colors to the
carapace. The common element between
the two is melanin-producing melanocytes,
but it is not known exactly which is induced
by what stimulation. A common theory
holds that they are controlled by the genes
(an intrinsic element)(Seiji, 1970; Oikawa,
1976; Prota, 1980). The effects of a ray of
light (Seiji et. al., 1973) or hormones
(melanocyte-stimulating, | adrenocortical,
sex, and thyrotropic hormones) (Seiji, 1970;
Parker, 1974; Snell, 1966) can provide the
necessary conditions for melanocytes to
stimulate the generation of melanosomes
(brightness of carapace). Empirically,
carapace patterns of the hawksbill are
bilaterally symmetrical. Since the way the
pattern starts out at individual scutes is
identical, it is presumed that those patterns
(two types of the melanin arrangement)
have more genetic (innate) factors than en-
vironmental (acquired) factors. However,
when a hawksbill turtle is raised without
sunlight, its carapace colors become lighter.
Thus, environmental factors are believed to
be involved when it comes to brightness.

In general the developmental habitat does
not seen to differ from that of the adult, and
juveniles and adults are taken together in
the same forging areas (Limpus, 1992;
Broderick et al., 1994). So those turtles,
within the same feeding habitat, were consi-
dered sub-adults or adults. In this study,

Current Herpetol. 19(2) 2000

no correlation was observed between C1W
and brightness (Fig. 1). This fact at least
indicates that in the case of a hawksbill with
a C1W of 18.3 cm (SCL=51.3 cm, calculat-
ed by SCL=4.3527 (C1 W)?-8484, r2=0.9529,
N=340, unpublished data) or greater there
is little change in scute color as it grows.
However, there have been reports of
changes in carapace patterns of other turtle
species due to sexual differences after
maturity (Barbour and Carr, 1940; Moll,
1980), as well as due to life stages and size
(McCoy, 1968; Balazs, 1986). Considering
those reports, there is a possibility that en-
dogenous substances would change the pat-
terns. Therefore, it may be worth studying
the changes in patterns of hawksbills before
and after sexual maturity or in relation to
the differences of the sexes.

Barnacles that I examined in this study
mostly belonged to the genus Chelonibia by
morphology, although I could not classify
them as to species. There is a report that
these same barnacles on the carapace are
typical of hawksbills on the Caribbean coast
of Costa Rica, but not of Chelonia (Chelo-
nia mydas) (Carr et al., 1966). Chelonibia,
the commensal barnacle on sea turtles, pos-
sesses marine qualities and is gregarious in
shallow water (on the coast). Elements that
regulate the distribution of Cirripedia in-
cluding barnacles are temperature and the
concentration of salt. Since the embryos
scatter seeds while swimming freely, the
durability against external conditions does
regulate the geographical distribution
(Shiino, 1964). With these facts about bar-
nacles in mind and the fact that the greater
the C1W the more significant the C1 with
barnacles is and that barnacles do not at-
tach to Cl with C1W of 20.3 cm
(SCL=56.0cm) or smaller (Fig. 2), it is
highly likely that turtles with barnacles have
different migration routes from turtles
without barnacles and these turtles are a set
of turtles which frequently use shallow
water (coast). The fact that Cls with at-
tached barnacles were significantly lower in

KOBAY ASHI—CARAPACE OF HAWKSBILL TURTLE 95

brightness (somewhat dark) than other Cls
is considered to be caused by the accumu-
lated melanin of exogenous elements (such
as the amount of sunlight) after the
hawksbills moved to shallow water. In
fact, all the species of sea turtles move at
least a small distance from the feeding
habitat to the breeding habitat. After mat-
ing, males return to the feeding habitat
while females move to the nesting habitat
(Limpus and Miller, 1993). After nesting
for several months, females return to the
feeding environment, preparing for the next
breeding (Limpus and Miller, 1993; Miller,
1985). Also, the feeding habitat is general-
ly the same whether they are baby turtles,
sub-adults, or adults (Limpus, 1992;
Broderick et al., 1994). In Cuban popula-
tions, sexual maturity is reached when SCL
is 51-55cm, the size of the nesting turtle
ranges from 60 to 85cm, and male sexual
maturity is reached when SCL is 68cm or
greater (Moncada et al., 1998). What those
facts suggest, and what has become clarified
in this paper is that a turtle with attached
barnacles will move to shallow water or the
coast for certain reasons (e.g., mating or
nesting) and sexual maturity and the differ-
ence between the sexes may be linked to the
attachment of barnacles. The hypothesis
fits the report that small (SCL<50 cm) log-
gerheads do not have these barnacles in the
Mediterranean sea (Gramentz, 1988).

I used 256 phases of shade to measure the
brightness of hawksbill carapaces. Empiri-
cally speaking, because the brightness of the
carapace is believed to include environmen-
tal elements, it is possible to gather ample
information by linking the brightness to
other environmental factors. In this study,
as an example, with the characteristics of
one turtle with attached barnacles I was able
to express the brightness and size. Espe-
cially, for marine turtles which are difficult
to follow and examine in the ocean with the
naked eye, it is possible to obtain data of
brightness of carapace on stranded turtles
or live turtles. Thus, this suggests that in-

formation based on the brightness can add
to a fuller understanding of the environ-
mental characteristics of hawksbill turtles.

ACKNOWLEDGEMENTS

I thank the staff of the Ministry of Fish-
ery in Cuba, Breading Center on Isla de Pi-
nos, and numerous fishermen of Nuevitas
and Doce Leguas for the field work. Fund-
ing was provided by the Japan Bekko As-
sociation. Also for contributing to the suc-
cess of this study, I am indebted to Mr. G.
Webb, Mr. C. Manolis (Wildlife Manage-
ment International Pty. Ltd.), and Mr. T.
Tubouchi (Japan Wildlife Research Cen-
ter).

LITERATURE CITED

BALAzs, G.H. 1986. Ontogenetic changes in
plastron pigmentation of hatchling Hawaiian
green turtle. J. Herpetol. 20: 280-282.

BARBOUR, T. AND A. F. Carr, Jr. 1940. An-
tillean terrapins. Mem. Mus. Comp. Zool. 54:
379-415.

BRODERICK, D., C. Moritz, J.D. MILLER, M.
GUINEA, P.I.T. PRINCE, AND C. J. LIMPUS.
1994. Genetic studies of the hawksbill turtle,
Eretmochelys imbricate: evidence for multiple
stocks in Australian waters. Pacific Conserv.
Biol. 1: 121-131.

Carr, A., H. HIRTH, AND L. OGREN. 1966.
The ecology and migrations of sea turtles, 6:
The hawksbill turtle in the Caribbean sea.
Amer. Mus. Novit. (2248): 1-29.

GRAMENTZ, D. 1988. Prevalent epibiont sites
on Caretta caretta in the Mediterranean sea.
Naturalista Sicil. Ser.4 12(1/2): 33-46.

Limpus, C. L. 1992. The hawksbill turtle, Erez-
mochelys imbricata, in Queensland: Popula-
tion structure within a Southern Great Barrier
Reef feeding grand. Wildl. Res. 19: 489-506.

Limpus, C. J. AND J. D. MILLER. 1993. Family
Cheloniidae. p. 1-113. Jn: C. J. Glasby, G. J.
Ross, and P. L. Beesley (eds.), Fauna of Aus-
tralia, Vol.2A, Amphibia and Reptilia. Aus-
tralian Government Publishing Service, Can-
berra, Australia.

MATSUURA I. AND K. NAKAMURA. 1993. At-
tachment Pattern of the Turtle Barnacle

Chelonibia testudinaria on Carapace of Nest-
ing Loggerhead Turtle Caretta caretta. Jpn.
Soc. Fish. Sci. 59: 1803.

McCoy, C.J. 1968. The development of
melanism in a Oklahoma population of
Chrysemys scripta elegans (Reptilia: Testudin-
idae). Proc. Oklahoma Acad. Sci. 47: 84-87.

MILLER, J.D. 1985. Embryology of marine
turtles. p. 1-269. In: C. Gans, F. Billett, and
P. F. A. Maderson (eds.), Biology of the Rep-
tilia, Vol. 14A. Wiley & Sons, New York.

MoNcApDA, F., E. CARRILLO., A. SAENZ., AND G.
NopARsE. 1998. Reproduction and Nesting
of the hawksbill turtle, Eretmochelys imbrica-
ta, in the Cuban Archipelago. Chelonia. Con-
serv. Biol. 3: 257-263.

Morr, E. O. 1980. Natural history of the river
terrapin, Batagur baska (Gray) in Malaysia
(Testudines: Emydidae). Malaysian J. Sci. 6:
23-62.

OIkKAWA, J. 1976. Melanocyte and Melanin.
Biochemistry 48: 872-888. (in Japanese)

PARKER, F. 1974. Skin and Hormones. p. 977-
993. In: R.H. Williams (ed.), Textbook of
Endocrinology, 5th edition. W. B. Saunders
Comp., Philadelphia.

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PROTA, G. 1980. Recent advances in the che-
mistry of melanogenesis in mammals. J. In-
vest. Dermatol. 75: 122-127.

SEI, M. 1970. Melanin. p. 292-357. In: K.
Okutani, K. Iwashita, and K. Takeuchi (eds.),
Biochemistry and Pathology of Skin.
Igakushoin, Tokyo. (in Japanese)

SEIJI, M., TAKAHASHI, M. AND Y. TANABASHI.
1973. Regulatory mechanisms of human skin
against light. p.75-92. In: M. Seiji, Y.
Takase, J. Toda, H. Miyazaki, and T.
Morikawa (eds.), Light and Skin. Kanehara
Shupan, Tokyo. (in Japanese)

SNELL, R.S. 1966. Hormonal of control of
pigmentation in man and other mammals.
p. 447-466. In: Advances in Biology of Skin:
The Pigmentary System, Vol. VIII. Pergamon
Press, Oxford, U.K.

SHIINO, H. 1964. Crustacean. p. 23-151. In: T.
Uchida (ed.), Systematic Zoology Vol.7:
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Japanese)

Accepted: 31 August 2000

Current Herpetology 19(2): 97-111., Dec. 2000
© 2000 by The Herpetological Society of Japan

Literature Survey on Predators of Snakes in Japan

Kon TANAKA* ANp AkrRA MORI

Department of Zoology, Graduate School of Science, Kyoto University, Sakyo-ku,

Kyoto 606-8502, JAPAN

Abstract: Characteristic defensive behaviors of snakes and their ecological
and morphological correlates have been well documented. Biological inter-
pretations of these characteristics, however, often suffer from a paucity of in-
formation on actual predators of snakes. Concerning natural predators of
Japanese snakes, neither quantitative data nor a systematic review are availa-
ble. Here, we review and synthesize the published accounts of predators of
snakes in Japan. We confirmed 59 species/subspecies of predators and 21
species of prey snakes. We hope that this review will stimulate biologists and
naturalists to record further predatory events on snakes and help clarify the

defensive mechanisms of snakes.

Key words:

INTRODUCTION

Prey-predator interaction is one of the
most important aspects of evolutionary bi-
ology, because this interaction can affect the
evolution of phenotypic features, such as
morphology and behavior, of both prey and
predators (Feder and Lauder, 1986).

Snakes, typical carnivorous animals, have
been well-studied as “predators”. Substan-
tial information on their diets, feeding be-
havior, and associated ecological and mor-
phological characteristics have been ac-
cumulated (Mushinsky, 1987; Greene,
1997). Attention has also been paid to
snakes as “prey”, and characteristic defen-
sive behaviors and ecological and morpho-
logical correlates have been documented
(Greene, 1988; Pough, 1988). However,

* Corresponding author. Tel: +81-75-753-
4075; Fax: +81-75-753-4113.

E-mail address: koji@ci.zool.kyoto-u.ac.jp
(K. Tanaka)

Snake; Predator; Prey; Review; Japan

biological interpretations of their defensive
mechanisms often suffer from a paucity of
information on actual predators of snakes.

In Japan, 38 species of snakes (excluding
sea snakes) are currently recognized (Hikida
and Sengoku, 2000). For several species of
them, food habits are well documented, and
Mori and Moriguchi (1988) reviewed the
published data on natural diets of Japanese
snakes. On the other hand, information on
predators of Japanese snakes are scattered,
and neither quantitative data nor a systemat-
ic review are available concerning natural
predators of snakes in Japan. We have at-
tempted to review and synthesize the pub-
lished accounts of predators of snakes in
Japan, and we provide a list of natural pre-
dators of Japanese snakes based on more
than 200 references.

We employed the following criteria for
citation. 1) The data used here were limited
to predatory events observed within Japan.
For the snakes that are not endemic to
Japan, predatory events observed in other

countries were not included. 2) Although
humans could play an important role in the
evolution of phenotypic features of snakes,
predation by Homo sapiens was not consi-
dered. 3) Predation under captive or ex-
perimental conditions was excluded. 4) Ac-
tual ingestion of the snake by the predator
was not considered. For example, observa-
tion of a flying bird carrying a snake or ob-
servation of a predatory attempt without
actual ingestion was considered as a preda-
tory event and included in the present cita-

TABLE 1.

Current Herpetol. 19(2) 2000

tion. 5) Predation obviously considered as
carcass feeding was excluded. However,
unless evidence of carcass feeding was avail-
able, predation on dead snakes was includ-
ed. 6) General descriptions of food habits
and speculated predators without concrete
evidences were excluded. 7) If the data ob-
viously obtained from the same survey were
published in separate papers, we cited only
a representative one. 8) Predators of sea
snakes were excluded, and we did not make
an extensive literature survey for sea snakes.

List of animals reported as predators of Japanese snakes. Subspecies names of preda-

tors are listed if identification is possible from the literature.

Class Order Species* Symbol
Arachnida Araneae Achaearanea tepidariorum Ar-1
Nephila maculata Ar-2
Osteichthyes Anguilliformes Anguillia marmorata O-1
Salmoniformes Oncorhynchus masou masou O-2
Oncorhynchus mykiss O-3
Salvelinus leucomaenis pluvius O-4
Salvelinus leucomaenis O-5
Amphibia Urodela Andrias japonicus A-1
Anura Bufo sp. A-2
Rana catesbeiana A-3
Rana nigromaculata A-4
Rana ornativentris A-5
Reptilia Squamata Agkistrodon blomhoffii R-1
Dinodon orientale R-2
Dinodon rufozonatum walli R-3
Dinodon semicarinatum R-4
Elaphe climacophora R-5
Elaphe quadrivirgata R-6
Hemibungarus japonicus boettgeri R-7
Hemibungarus japonicus japonicus R-8
Hemibungarus macclellandi iwasakii R-9
Ovophis okinavensis R-10
Trimeresurus elegans R-11
Trimeresurus flavoviridis R-12
Aves Ciconiiformes Ardea purpurea manilensis V-1

Nycticorax nycticorax nycticorax V-2

TANAKA & MORI—PREDATORS OF JAPANESE SNAKES 99

TABLE 1. Cotinued.

Falconiformes Accipiter gentilis fujiyamae V-3
Aquila chrysaetos japonica V-4
Butastur indicus V-5
Buteo buteo japonicus V-6
Circus spilonotus spilonotus V-7
Falco tinnunculus interstinctus V-8
Haliaeetus albicilla albicilla V-9
Milvus migrans lineatus V-10
Pernis apivorus orientalis V-11
Spilornis cheela perplexus V-12
Spizaetus nipalensis orientalis V-13
Galliformes Phasianus colchicus V-14
Phasianus soemmerringii V-15
Strigiformes Strix uralensis V-16
Coraciiformes Halcyon coromanda major V-17
Passeriformes Corvus corone orientalis V-18
Corvus macrorhynchos japonensis V-19
Corvus sp. V-20
Lanius bucephalus bucephalus V-21
Sturnus cineraceus V-22
Mammalia Primates Macaca fuscata yakui M-1
Carnivora Felis bengalensis euptilura M-2
Felis catus M-3
Felis iriomotensis M-4
Herpestes javanicus M-5
Meles meles anakuma M-6
Mustela itatsi M-7
Mustela sibirica coreana M-8
Mustela vison M-9
Nyctereutes procyonoides viverrinus M-10
Ursus arctos M-11
Vulpes vulpes japonica M-12
Vulpes vulpes schrencki M-13
Unidentified species** M-14
Artiodactyla Sus scrofa riukiuanus M-15

* Scientific names follow literature listed below. Thus, they are not necessarily consistent with
those used in the original literature.
Arachnida, Osteichthyes, and Mammalia: Hidaka, T. (ed.) 1998. The Encyclopaedia of Animals
in Japan, Extra Volume, General Index and Red Lists of Threatened Animals in Japan.
Heibonsha, Tokyo. 334 p.
Aves: The Ornithological Society of Japan, Editorial Committee for List of Japanese Birds.
1997. List of Japanese Birds. Jpn. J. Ornithol. 46(1): 59-91.
Amphibia and Reptilia: Hikida, T. and S. Sengoku. 2000. Japanese common names of amphib-
ians and reptiles: past and present. Bull. Herpetol. Soc. Jpn. 2000(1): 20-33.

** Authors speculated that the predator was Canis familiaris or Felis catus based on the feces.

100 Current Herpetol. 19(2) 2000

literature without a original English title
and/or English name of the periodical.
For the convenience of Japanese readers, a
list of the literature in Japanese and the ta-

However, a few items of information ob-
tained during our general survey are
presented as an appendix.

We attempted to give translations for

TABLE 2. List of literature records for predators of Japanese snakes. See Table 1 and Literature
List for symbols of predators and reference No., respectively.

Snake species

Achalinus spinalis

Achalinus werneri
Agkistrodon blomhoffii

Amphiesma pryeri pryeri
Amphiesma vibakari vibakari
Cyclophiops herminae

Cyclophiops semicarinatus

Dinodon orientale

Dinodon rufozonatum rufozonatum
Dinodon rufozonatum walli
Dinodon semicarinatum

Elaphe climacophora

Elaphe conspicillata

Elaphe quadrivirgata

Predator (reference No.)

R-1 (23, 76, 103, 104, 131, 197), R-2 (188, 198), R-6 (44,
94, 109, 186)

M-3 (187)

R-4 (171), R-12 (74, 75, 97)

O-3 (120)

A-2 (176)

R-6 (6, 92, 165, 175, 185, 191)

V-4 (39, 48, 154*), V-5 (3, 56, 82), V-20 (161)

M-7 (16, 119)

R-4 (96), R-10 (73, 95), R-12 (70, 71, 73, 75, 97)

M-7 (224), M-14 (210)

R-1 (54, 98, 99), R-2 (147), R-6 (43, 54, 94)

V-5 (55**), V-21 (85)

V-12 (181)

M-4 (136, 137, 138, 205, 218)

R-4 (12, 96, 113, 169), R-10 (95), R-12 (69, 70, 71, 73,
15, 97.101)

V-1 (193), V-5 (110)

M-S5 (2)

R-6 (127)

M-3 (32)

M-2 (47, 146)

M-4 (136, 137, 138, 139, 205, 218)

Ar-2 (7)

R-4 (96), R-10 (95, 182), R-12 (73, 75, 97)

M-5 (195)

Ar-1 (8)

A-1 (143), A-4 (151), A-5 (67)

R-5 (178, 192), R-6 (25, 44, 197)

V-3 (64), V-4 (9, 39, 48, 144, 154*, 155, 157, 160, 163,
164, 174, 180, 183, 204, 209, 212, 214), V-5 (3, 55**, 82,
123), V-6 (59, 211, 221), V-9 (31, 108), V-10 (57), V-13
(19, 48, 111, 112, 200, 206), V-14 (226), V-16 (14), V-19
(84), V-21 (18, 107)

M-2 (47, 213), M-3 (116, 213), M-7 (179), M-11 (66)
R-6 (190)

V-4 (39, 154*), V-5 (3, 123), V-6 (56, 162), V-11 (1),
V-13 (200), V-17 (42), V-21 (18)

A-3 (33)

TANAKA & MORI—PREDATORS OF JAPANESE SNAKES 101

TABLE 2. Cotinued.

Elaphe taeniura schmackeri
Elaphe sp.

Hemibungarus japonicus japonicus
Ovophis okinavensis

Ramphotyphlops braminus

Rhabdophis tigrinus tigrinus

Trimeresurus elegans

Trimeresurus flavoviridis

Trimeresurus sp.
Unidentified snake (Colubridae)

Unidentified snake

V-3 (64), V-4 (39, 144, 154*, 163, 164, 174, 209, 214),
V-5 (3, 86**, 87, 105, 159), V-6 (36, 52, 221), V-10 (63,
117, 159, 172), V-13 (48, 61, 200), V-21 (58, 106, 114)
M-1 (91)

M-4 (136, 137, 138, 139, 205)

V-13 (111)

R-12 (75)

R-4 (96), R-12 (72, 75, 97, 171, 182)

M-S5 (195)

R-3 (49), R-4 (12), R-7 (170), R-8 (171), R-9 (171)

M-S5 (2, 189)

O-4 (29)

Aa Cah

R-6 (13, 22, 44, 94, 145)

V-3 (64), V-4 (39, 154*, 180, 209), V-5 (62, 89, 132),
V-6 (36, 51, 52), V-11 (1,159), V-18 (37), V-21 (18, 85,
106, 133, 134, 140, 152, 153, 211), V-22 (194)

M-10 (35)

O-1 (220***)

R-3 (168, 171)

V-5 (167), V-12 (24, 88, 101, 149, 181, 223)

M-4 (136, 138, 205)

R-4 (96, 169, 171), R-10 (118), R-12 (72, 75, 97)

M-5 (207)

M-15 (219)

V-7 (51)

M-2 (166), M-4 (136, 137, 205)

O-2 (141), O-5 (199)

A-2 (176), A-4 (65)

R-4 (44), R-6 (30), R-11 (11, 12), R-12 (10, 12)

V-2 (93), V-4 (4, 9, 34, 39, 48, 79, 115, 121, 122, 126,
148, 154*,155, 156, 157, 158, 164, 174, 180, 183, 201,
204, 208, 212, 215, 216), V-5 (15, 41, 77, 78, 82, 83, 130,
185, 196), V-6 (36, 52, 60, 88, 102, 162), V-8 (203),
V-10 (50), V-11 (112), V-12 (24, 40, 101, 128), V-13 (20, 21,
48, 53, 80, 81, 100, 124, 135, 216), V-15 (5), V-17 (150),
V-18 (217), V-19 (90), V-20 (26), V-21 (27, 85, 106, 129,
2902225)

M-2 (47, 177, 213), M-4 (45, 139, 167), M-5 (2, 28, 46,
68, 125), M-6 (17), M-8 (142), M-9 (202), M-12 (184),
M-13 (38, 173), M-14 (210), M-15 (219)

* This paper seems to include both original and cited data. Because it was not possible to distin-

guish them, we cited all of the data.

** Jdentification of the prey species is based on personal communication from the author.
*** The author stated that he had seen a record of an Anguillia marmorata that had swallowed a

Trimeresurus elegans.

102

bles with Japanese common names will be
published in the Bulletin of the Herpetolog-
ical Society of Japan.

Predators of Japanese snakes revealed by
the literature survey are listed in Table 1.
Predators of each species or subspecies of
snakes along with references are listed in
Table 2. We confirmed 59 species/subspe-
cies of predators, encompassing a range
from invertebrate arachnids to large mam-
mals. At least one predator was recorded
for 21 species of snakes, which is approxi-
mately half of the Japanese terrestrial
snakes. It should be noted that these tables
are not to be considered an all-inclusive
summary of the predators of Japanese
snakes. We hope that this review will
stimulate biologists and naturalists to ob-
serve and report further predatory events on
snakes and promote future studies on
defensive mechanisms of snakes.

ACKNOWLEDGMENTS

Our special thanks are due to A. Azuma,
T. Deguchi, M. Hasegawa, T. Hikida, T.
Hirai, N. Ichikawa, M. Imafuku, M. Izawa,
S. Kadowaki, K. Karasawa, S. Kubokami,
H. Kuroda, T. Maenosono, H. Matsubara,
R. Miura, M. Motokawa, H. Moriguchi, N.
Narumi, M. Nishimura, Y. Nitani, T. Oi, S.
Okada, H. Okawa, G. Ogura, I. Ono, N.
Sakaguchi, K. Sato, S. Suginohara, S.
Tachibana, K. Taguchi, S. Takenaka, M.
Tatara, T. Tochimoto, Ma. Toda, Y. Tom-
ida, M. Toyama, R. Tsushima, K. Ueda, T.
Wada, S. Watanabe, Yo. Yamada, S.
Yamagishi, and M. Yoshida for providing
literature and/or personal information on
predators of snakes, and to the Research
Center, Wild Bird Society Japan (Y. Kanai,
R. Kurosawa, Ya. Yamada, K. Fukui, and
N. Kato) and the Yamashina Institute for
Ornithology for providing literature. We
also thank K. Araya, M. Fujioka, R.C.
Goris, T. Hayashi, K. Hidaka, H. Higuchi,
T. Izumi, M. Kajita, K. Kawamura, G.
Masunaga, S. Mori, O. Murakami, Y.

Current Herpetol. 19(2) 2000

Muroyama, N. Nakagawa, H. Ota, Y.
Sawara, T. Taniuchi, and Mi. Toda for
their comments on our preliminary ques-
tionnaire for this study.

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on Iriomote-jima Island. Unpublished

206.

207.

208.

209.

210.

7
Pm Pap

213.

214.

Zils

216.

Current Herpetol. 19(2) 2000

Bachelor of Science Thesis, Ryukyu
Univ., Okinawa. 28 p. + 9 tabs + 13 figs
+ 3 pls. (**)

WATANABE, T. 1985. Ecological status
of the golden eagle and Hodgson’s hawk-
eagle in the Higashi-kamahara region.
Wild Birds of Niigata 62: 4-6. (**)
WATASE, S. 1911. Mongooses reproduc-
ing on Tonaki-jima Island. Zool. Mag.
23: 109-110. (**)

Wild Bird Society of Hyogo Golden Eagle
Group. 1978. The golden eagle: Birds
and gliders soaring in an ascending cur-
rent. Searching for the golden eagle in the
Chugoku mountains, Japan. Anima 6(2):
21-27. (**)

Wild Bird Society of Japan. 1976. The
golden eagle. p. 167-223. In: Research on
Designated Birds, Contract Research of
Environment Agency, 1975. Environment
Agency, Tokyo. (**)

Wild Bird Society of Japan, Yanbaru
Chapter. 1997. Reports on the Distribu-
tion of Endangered Animals and In-
troduced Animals in the Northern Part of
Okinawajima Island and Influence of In-
troduced Animals Upon Endangered
Animals. Wild Bird Society of Japan,
Yanbaru Chapter, Okinawa. 86p. + 5
apps. (**)

YAMAGISHI, S. Personal Communication.
Yamamoto, Y. 1997. After the Golden
Eagle. Kobe Shimbun Publ. Center,
Nojigiku Bunko, Kobe. 275 p. (**)
YAMAMURA, T. 1996. An Encyclopedia
of the Tsushima Leopard Cat. Data
House, Tokyo. 78 p. (**)

YAMANOI, A. AND I. KATAYAMA. 1983.
Food habits of the golden eagle in the
Joshin-etsu region. (abstract) p. 265. In:
Proceedings of the 30th Annual Meeting
of the Ecological Society of Japan. The
30th Annual Meeting of the Ecological
Society of Japan Preparatory Committee,
Faculty of Science, Shinshu Univ., Mat-
sumoto. (**)

YAMAZAKI, T. 1985. Food habits and
foraging behavior of the golden eagle in
the Suzuka mountains. (abstract) Tori
34(2/3): 83. (**)

YAMAZAKI, T. 1989. Distribution pat-
terns of the golden eagle and Hodgson’s
hawk-eagle in the Suzuka mountains. (ab-
stract) Aquila chrysaetos 7: 37. (**)

TANAKA & MORI—PREDATORS OF JAPANESE SNAKES fii

aT.

Z18.

219.

220.

Zo.

223.

224.

ググ 9

YANAGIMACHI, K. 2000. Carrion crows
struggling to handle a snake. Birder 14(1):
65. (**)

YAsUMA, S. 1982. Ryukyu Islands. Enig-
matic Formation Suggested by Geograph-
ic Distributions of Animals. Tokai Univ.
Press, Tokyo. 208 p. (**)

Yasuma, S. 1983. Ryukyu wild boars on
Iriomote Island. Animals and Zoos 35(1):
12-15. (**)

YoxotTsukA, M. 1994. After the Irio-
mote Cat. Takarajimasha, Tokyo. 237 p.
C*)

YONEKAWA, H., M. KAWABE, AND Y.
IwAMI. 1995. The breeding biology of
common buzzard, Buteo buteo and fac-
tors in decreasing their breeding popula-
tion in the Tokachi plain, Hokkaido. Bull.
Higashi Taisetsu Mus. Nat. Hist. 17: 1-
ar)

YosHIDA, T. 1952. Notes on animals im-
paled by the shrike. Science through Ex-
periments 3(4): 320-323. (**)

YoOsHIMI, K. 1991. The crested serpent
eagle on Iriomote-jima Island. Nat. Hist.
Jpn. 5(3): 4-11. (**)

YotsumMoTo, T. 1959. Mink grazing to
the island of Amami Oshima, Ryukyu.
Japan Wildlife Bulletin 17(1): 156-158.
30

YUASA, S. 1976. Five questions about
animals impaled by shrikes. p. 211-214.
In: T. Ueki (ed.), Animals of Toyama
Prefecture. Our Loving Friends 6, Toya-

ma Bunko. Kogen Shuppan, Toyama.

SUPPLEMENT

226. M. HaAsEGAWA. Personal Communica-
tion.

227. TocHIMoToO, T. AND K. SuHimizu. 2000.
On the foods of the Japanese giant
salamander Andrias japonicus (Tem-
minck, 1837). (Oral presentation at the
39th Annual Meeting of the Herpetologi-
cal Society of Japan)

APPENDIX

Predators of sea snakes
Predator—Chondrichthyes, Carcharhiniformes :
Galeocerdo cuvier
Prey—Emydocephalus ijimae (App. 2); Hydro-
Dhis melanocephalus (App. 2); Laticauda
semifasciata (App. 2); Pelamis platurus (App.
1); Unidentified sea snake (App. 2)

Literature

App. 1. Ucunipa, K. 1979. An Essay on the
Natural History of Fishes. Rippu Shobo,
Tokyo. 242 p. (**)

App. 2. YANO, K. 1998. Sharks. Mysterious
Ecology of the Chondrichthyes. Tokai Univ.
Press, Tokyo. 223 p. (**)

Accepted: 31 August 2000

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113

Titles of Papers Presented at the 39th Annual Meeting of the
Herpetological Society of Japan (4-5 November 2000
at Ryukyu University, Okinawa)

Abstracts in Japanese will appear in
Bulletin of the Herpetological Society of

Japan, Vol. 2001, No. 1.

[Turtles]

1 。

Satellite tracking of male loggerhead tur-
tle (Caretta caretta) migration. By:
Naoki Kamezaki, Kojiro Mizuno, Masato
Kobayashi, and Takaaki Nishi.
Distribution of nesting sites of sea turtles
in the Okinawa Islands, Miyako Islands,
and Yaeyama Islands. By: Koichi
Hirate, Naoki Kamezaki, Akira Kikuka-
wa, Tetsuya Kondo, Kenji Kuroyanagi,
Keiichi Nomura, and Tatsuya Shima.

. Seasonal changes of movement pattern in

the yellow-margined box turtle
(Cistoclemmys flavomarginata evelynae).
By: Koreyuki Kurosawa and Masako Iza-
wa.

Phylogenetic relationships of the genus
Mauremys (Testudines: Bataguridae).
By: Masanao Honda, Yuichirou Yasuka-
wa, and Hidetoshi Ota.

. Taxonomic status of two Pleistocene fos-

sil turtles, Cuora miyatai and Clemmys
yabei (Reptilia: Bataguridae), from
Kuzuu, Tochigi Prefecture. By:
Yuichirou Yasukawa and Ren Hirayama.
Sexual dimorphism in the Chinese soft-
shelled turtle, Pelodiscus sinensis. By:
Hiroyuki Sato, Atsushi Kaneko, and
Hidetoshi Ota.

. Normal embryonic development of the

Chinese soft-shelled turtle, Pelodiscus

sinensis. By: Masayoshi Tokita and
Shigeru Kuratani.
Distribution, food habitats, and

reproductive cycles of four introduced
freshwater turtles (Trachemys scripta ele-

gans, Mauremys mutica, Chinemys
reevesii, and Pelodiscus sinensis) on
Okinawa Island. By: Dai Hatanaka and
Takeshi Sasaki.

[Lizards]

10;

Oke

13%

14.

15.

16.

Phylogeny and biogeography of the
agamid genus Japalura in East Asia. By:
Hidetoshi Ota, Masanao Honda, Szu-
Lung Chen, and Tsutomu Hikida.
Sexual size dimorphism of the agamid liz-
ard Japalura polygonata. By: Satoshi
Tanaka.

Distribution of the genus Gekko in
Nagasaki Prefecture. By: Takanori Mat-
suo.

Phylogenetic relationships among the
Japanese members of the genus Gekko.
By: Mamoru Toda, Hidetoshi Ota, and
Tsutomu Hikida.

Distribution of house geckos in Taiwan
and Amamioshima Island of the Ryukyu
Archipelago. By: Takahiko Kuze and
Hidetoshi Ota.

Clonal composition of the parthenogenet-
ic gecko, Lepidodactylus lugubris, in Tai-
wan. By: Saiko Yamashiro, Wen-hao
Chou, and Hidetoshi Ota.

Taxonomic notes on specimens of the ge-
nus Scincella (Squamata: Scincidae) from
Vietnam. By: Szu-Lung Chen, Nikolai
L. Orlov, Ilya Darevsky, Hidetoshi Ota,
and Tsutomu Hikida.

Three undescribed species of the flat-bo-
died water skinks of the genus
Tropidophorus (Reptilia: Scincidae) from
Thailand and Vietnam. By: Tsutomu
Hikida, Hidetoshi Ota, Jarujin Nabhitab-
hata, and Nikolai L. Orlov.

Additional notes on reproduction of

114

Takydromus lizards of the Nansei Shoto.
By: Sen ‘Takenaka and _ Hajime
Moriguchi.

[Snakes]

18.

19.

20.

23%

24.

26.

Geographic morphological variation in a
colubrid snake, Dinodon semicarinatum,
from the central Ryukyus, Japan. By:
Isao Takiguchi and Hidetoshi Ota.
Appearance rate of Elaphe quadrivirgata
in an outdoor enclosure. By: Hajime
Moriguchi.

Geographic variations in morphology of
the two Emydocephalus species, E. ijimae
and E. annulatus. By: Gen Masunaga
and Hidetoshi Ota.

Food habits of sea snakes in the Ryukyu
Archipelago. By: Mayuko Morishita,
Gen Masunaga, and Hidetoshi Ota.
Geographic variation and population sys-
tematics Of Trimeresurus flavoviridis and
T. tokarensis in the central Ryukyus,
Japan. By: Hiroko Nagatomo and
Hidetoshi Ota.

Distribution and ecology of Trimeresurus
mucrosquamatus established on Okinawa
Island. By: Masahiko Nishimura and
Hiroyuki Akamine.

A case of envenomation by the habu,
Trimeresurus flavoviridis. By: Setsuko
Iwanaga, Mamoru Toda, Tadafumi
Maenosono, and Hidetoshi Ota.
Classification of A gkistrodon (sensu lato)
and 7Jrimeresurus (sensu lato) based on
micro-ornamentation of scale surface.
By: Michihisa Toriba.

Literature survey on predators of snakes
in Japan. By: Koji Tanaka and Akira
Mori

[Reptiles general]

General growth model and its examina-
tion by reptiles’ growth data. By: Masa-
mi Hinoue.

[Salamanders]

28.

On the prey of the Japanese giant
salamander Andrias japonicus (Tem-

29:

30.

31.

32:

33.

34.

35.

36.

minck, 1837). By: Takeyoshi Tochimoto
and Kunikazu Shimizu.

Population structure of the Japanese gi-
ant salamander, Andrias japonicus, in
two streams of the Kannose River,
Hiroshima Prefecture. By: Sumio Oka-
da, Taeko Utsunomiya, Yasuaki Utsu-
nomiya, and Jun-ichi Naito.

Site selection for breeding in lotic water-
bodies by Hynobius tokyoensis. By: Sa-
dao Ihara.

Winter and spring breeding in same
population of Hynobius lichenatus. By:
Hiroshi Ota.

Sex chromosomes in Hynobius
nigrescens. By: Chikako Ikebe and Sei-
ichi Kohno.

Ontogenetic development of hind limbs
in Hynobius kimurae. By: Yasuchika
Misawa and Masafumi Matsui.

Variation in external morphology of
Hynobius naevius from Amagi City,
Fukuoka Prefecture. By: Atsushi
Tominaga, Masafumi Matsui, and Kanto
Nishikawa.

Genetic variation among populations of
Hynobius boulengeri. By: Kanto
Nishikawa, Masafumi Matsui, Sin’ichi
Sato.

Ecological comparison of two species of
salamander distributed in Hokkaido,
Japan. By: Takanori Sato, Shigehiro
Nakabayashi, Nobuyuki Narumi, Takehi-
to Ueda, and Yukihiro Kohmatsu.

[Frogs]

38.

39.

Differences in body size, growth, and age
at maturity between two populations of
the Japanese toad (Bufo bufo formosus).
By: Ayako Ochi.

Influence on spawning sites of the
Japanese Toads, Bufo torrenticola and B.
bufo formosus, by a flood of the Nozumi
River, Toyama Prefecture. By: Hisao
Nambu, Tamotsu Fukuda, and Yukima-
sa Araki.

Molecular phylogeny of the Eurasian
Bufo bufo group and allied groups. By:

40.

41.

42.

43.

44.

45.

46.

47.

48.

49.

50.

Wanzhao Liu and Masafumi Matsui.
€-Crystallin expresses in the Japanese tree
frog (Hyla japonica) lens as a taxon-
specific crystallin. By: Hisashi Kimoto,
Yutaka Fujii, Yuichirou Yasukawa,
Keiko Ishikawa, Kikuko Watanabe,
Akira Taketo, and Noboru Nakai.
Genetic variation among populations of
Rana pirica. By: Giedrius Trakimas,
Masafumi Matsui, Kanto Nishikawa, and
Kiyoshi Kasugai.

Allometry of Rana tagoi from Shishigata-
ni, Kyoto. By: Tadafumi Sugihara and
Masafumi Matsui.

Phylogenetic relationships among the
brown frogs from Eurasia. By: Tomoko
Tanaka-Ueno and Masafumi Matsui.

A preliminary experiment on recognizing
individuals of Rana nigromaculata. By:
Takeshi Kameyama, Toshihiro Morita,
and Sumio Okada.

Age determination of the Daruma pond
frog, Rana porosa_ brevipoda, by
skeletochronology. By: Wichase Khon-
sue, Masafumi Matsui, and Yasuchika
Misawa.

Notes on hibernation of a Japanese pond
frog, Rana porosa brevipoda. By: Tosi-
hiro Morita, Takeshi Kameyama, and Su-
mio Okada.

Examples of homing ability of Rana
catesbeiana. By: Junsuke Marunouchi,
Tamotsu Kusano, and Masayuki Sumida.
Breeding of the ranid frog, Rana narina,
on Okinawajima Island, Ryukyu Ar-
chipelago. By: Masanao Toyama.
Identity of Rana okinavana. By:
Masafumi Matsui.

Discovery of the rice frog Rana lim-
nocharis from the area along the
Watarase River in the northern part of
the Kanto District, Japan. By: Terutake

。

a2)

58%:

54.

356

56.

58.

59:

60.

115

Hayashi, Yuki Kimura, Kinen Akaba,
and Riichi Ishizuka.

The activity of male Rhacophorus
schlegelii breeding in paddy fields and a
technique for their capture. By: Satoshi
Osawa and Takehiko Katsuno.
Metapopulation dynamics of the
treefrog, Rhacophorus arboreus, in the
Kanazawa Castle grounds. By: Mitsu-
hiko Toda.

Effects of different diet conditions on
growth and development of Rhacophorus
arboreus tadpoles. By: Kenji Yoshinaga
and Takashi Kagaya.

Hibernation of stream-dwelling frogs.
By: Yoshinori Ogura.

Ontogenetic and taxonomic variation in
the speed with which anuran larvae react
to a startling stimulus. By: Masamichi
Yamashita, Harutake Kayamori, Tomio
Naitoh, and Richard J. Wassersug.

[Amphibians general]

Geographic and seasonal variations in the
species composition and body size of am-
phibians found in the feces of the Irio-
mote cat, Felis iriomotensis. By:
Shinichi Watanabe and Masako Izawa.
Current status and distribution of am-
phibians in South Korea. By: Jaeyoung
Song and Kyu-Hoi Chung.

Studies on in-situ conservation methods
for amphibians in Korea. By: Jaeyoung
Song and Jae-Han Shim.
Frog-dependent predation by the giant
water bug Lethocerus deyrollei (Hemip-
tera: Belostomatidae). By: Toshiaki
Hirai and Kazumasa Hidaka.

[General]
An example of DDT accumulation in am-
phibians and reptiles. By: Isamu Okochi

116

A New Book from the Society for the Study of Amphibians and Reptiles

AT THE UNIVERSITY OF KANSAS IN PICTURES
AND CONVERSATIONS by Sally Haines

Eighty-four Illustrations in Full Color

HIS BOOK IS IN THE FORM OF A GALLERY GUIDE TO A REMARKABLE EXHIBIT OF ILLUSTRATED HERPETOLOGICAL BOOKS DATING
] the 16th to the 20th centuries. It results from a display arranged by the Kenneth Spencer Research Library of the University of
Kansas on the occasion of the 1996 SSAR meeting held at the university. Books such as these, many of which have exquisitely handcolored
illustrations by some of the greatest natural history artists, are today kept under lock and key in institutional libraries and therefore are
scarcely known to most herpetologists, but the enthusiasm of the SSAR conference goers for the Kansas exhibit has encouraged us to
make this printed version available. The author, Sally Haines, Associate Special Collections Librarian at the Spencer Library, is in charge
of the natural history collections.

The bulk of this volume consists of full-color reproductions of illustrations from classic works together with extensive captions containing
information about the authors, the artists, and the books. It is an iconography of herpetology and an essential part of the history of the field.

Organization of the Book

Myth versus Reality: the Challenge of Illustrating Amphibians and Reptiles, by Kraig Adler
Part I — Herpetology to 1758: The Pre-Linnaean Period

Part II — Herpetology after 1758: Beyond Linnaeus

Part III — West of Eden: New World Herps

Part IV — The Seven Deadly Sins and other Evils: Herps as Symbol

References to Historical and Biographical Works

List of All Illustrated Herpetological Works in the Spencer Library
Chronological Index to Illustrated Herpetological Works in the Spencer Library

Some of the earliest books represented in this volume are by Pliny, Abbatius, Topsell, Seba, Linnaeus, and Résel von Rosenhof, with later
ones by Daudin, Riippell, Bonaparte, Schlegel, and Fayrer, among many others. There is a special section on books about the Western
Hemisphere, including works by Sloane, Catesby, Bartram, Spix, Holbrook, Cuvier, and Agassiz. The final section, on herps as symbol,
includes religious manuscripts, sea serpents, astronomical atlases, and even illustrations from the works of John Milton, Mark Twain,
and Walt Kelly. Included is a detailed listing of all illustrated herpetological works in the Spencer Library, which holds one of the great
collections of natural history books. This volume represents a veritable treasure trove of some of the finest illustrations of amphibians and
reptiles ever produced in book form and extensive bibliographic information about them.

Specifications: 190 pages, 8Y2 x 11 inches (21.5 x 28 cm), full color throughout (84 illustrations). Softcover. ISBN 0-916984-53-2.
To be published December 2000.

PRICES: Pre-publication price to SSAR members US$50; Institutions and non-members US$60.
SHIPPING COST: USA address, add US$3; non-USA address, add US$6

SEND ORDERS TO: Dr. Robert D. Aldridge, SSAR Publications Secretary, Department of Biology, Saint Louis University, 3507
Laclede Avenue, Saint Louis, Missouri 63103-2010, USA (telephone: area code 314, 977-3916 or -1710; fax: 314, 977-3658; e-mail:
ssar@slu.edu). Please make checks payable to “SSAR.” Overseas orders must be paid in USA funds using a draft drawn on American banks
or by International Money Order. Orders may also be charged to MasterCard or VISA (please provide the account number and card
expiration date). SSAR membership information and a complete list of all Society publications can be obtained on request to Dr. Aldridge.

A New Book from the Society for the Study of Amphibians and Reptiles

THE HERPETOFAUNA OF NEW CALEDONIA

by AARON M. BAUER and Ross A. SADLIER
with French summaries by Ivan Ineich and 189 color photographs

W CALEDONIA, INCLUDING THE LOYALTY ISLANDS

and an associated group of smaller islands and reefs,
is a French territory located in the tropical Southwest
Pacific equidistant from New Guinea, New Zealand,
and Australia. This ancient group of islands supports
one of the most highly endemic and species-rich
herpetofaunas in the Pacific region. Among the 71 spe-
cies of terrestrial reptiles, 86% are endemics, and most
belong to endemic genera. Despite being only 2.5% the
size of New Guinea, New Caledonia has 36% as many
lizard species. In addition, the New Caledonian barrier
reef system, one of the largest and most diverse in the
world, is inhabited by a dozen species of seasnakes. Be-
cause of the diversity of its flora and fauna and the fragil-
ity of its habitats, New Caledonia is regarded as a
biodiversity “hot spot,” one of the earth’s biologically

richest and most endangered terrestrial ecoregions.

This book—at the same time a scienufic monograph
and a field guide—is the first modern review of the
amphibians and reptiles of New Caledonia. It covers
the frogs, family Hylidae (1 species), geckos of the fami-
lies Diplodactylidae (20) and Gekkonidae (6), the
Scincidae (42), snakes of the families Boidae (1), Elapi-
dae (12, all marine), and Typhlopidae (2), and the sea
turtles, Cheloniidae (3). Geckos and skinks, in fact,
are the most numerous and dominant terrestrial verte-
brates in New Caledonia. These two groups have under-
gone extensive generic and specific diversification, in-
cluding the world’s largest living geckos (Rhacodactylus)
and more than a dozen genera of skinks including the

giant skinks (Phoboscincus).

The authors, Aaron M. Bauer (USA) and Ross A.
Sadlier (Australia) are both noted authorities on the
Pacific herpetofauna. Their extensive field work in New
Caledonia began more than 20 years ago. As a result of
their research, numerous new genera and species of New
Caledonian geckos and skinks have been described and
named, but this book represents the first synthesis of
their 20 years of study.

Organization of the Book

¢ Geography, Vegetation, Geological History, and
Biogeography

¢ Ecological Patterns of Terrestrial and Marine Spe-
Cles

¢ Conservation

¢ Humans and the Herpetofauna, including a History
of Scientific Studies

¢ Systematic Accounts of Genera and Species:
Keys; Synonymies; Descriptions; Distribution (spot
map for each species); Natural History; Conserva-
tion Status; Remarks and Status

¢ Incidental Taxa, Taxa of Questionable Occurrence,
and Those Erroneously Recorded from New Cale-
donia

¢ Fossil and Subfossil Species

* Literature Cited and Bibliography of New Caledo-
nian Herpetology (more than 1000 references)

* Gazetteer of Place Names; Index to Scientific Names

The 153 color photographs of animals depict nearly
every species. There are also 36 photographs of New
Caledonian habitats.

Specifications: 325 pages, 7 x 10 inches (18 x 22.5 cm), plus 189 color photographs of animals and habitats on 24
plates, 47 maps, 63 figures, and 4 tables. Clothbound. ISBN: 0-916984—55-9. To be published December 2000.

PRICES: Pre-publication price to SSAR members US$50; Institutions and non-members US$60.
SHIPPING: USA address, add US$3; non-USA address, add US$6.

SEND ORDERS TO: Dr. Robert D. Aldridge, SSAR Publications Secretary, Department of Biology, Saint Louis
University, 3507 Laclede Avenue, Saint Louis, Missouri 63103-2010, USA (telephone: area code 314, 977-3916
or—1710; fax:314, 977-3658; e-mail:ssar@slu.edu). Please make checks payable to “SSAR.” Overseas orders must
be paid in USA funds using a draft drawn on American banks or by International Money Order. Orders may also
be charged to MasterCard or VISA (please provide the account number and card expiration date). SSAR
membership information and a complete list of all Society publications can be obtained on request to Dr. Aldridge.

118

We are happy to inform you that the Fourth World Congress of Herpetology will be
held from 2nd to 9th December 2001, in the BMICH, Colombo Sri Lanka. This is the
first significant herpetology event of the new millennium and the organizing committee
is determined to make this the most memorable herpetological forum of the century.

The Registration—Full participant US $ 350, Students US $ 250, and Accompany
persons US $ 200. A surcharge of US $ 100 will be applied after 1st September 2001.

Deadline for abstracts—31 August 2001.

Registration entitlements for delegates—In addition to attendance at all formal ses-
sions, congress registration will be entitle you to:

Airport transfers (both ways)

Daily shuttle service from hotel to congress venue

Conference kit, which includes: Label badge, Special 4WCH pen, Special 4WCH note
pad, 4WCH picture post cards, Sri Lanka herp stickers, Welcome cocktail, Two coffee
breaks with snacks on each day of the congress, Lunch on all congress days, Congress
banquet, Full day professional excursion, Abstract book and the programme and many
more.

We have reserved special discount rate in leading five star hotel — shared room with
breakfast is only US $ 35 on wards.

We will be mailing you the congress Brochure and Registrations forms.

Sri Lanka is a herpetological hotspot in the world. In addition to upward 150 species
of amphibians, Sri Lanka is also home to diverse reptiles: Five species of marine turtles,
chelonians, crocodiles, varanids, agamids, skinks, lacertids, chameleons, geckos, and
snakes (from primitive uropeltids to modern vipers) inhabit our small island of 64,742
square km in area.

I look forward to welcoming you in Sri Lanka in December 2001.

Pre-registration & information:

4WCH Promotions Office,
95 Cotta Road,

Colombo 8,

SRI LANKA.

E-mail: admin@4wch.com
Internet: http://www.4wch.com

Congress organizer/director:

Anslem de Silva,

Faculty of Medicine,
University of Peradeniya,
Peradeniya,

SRI LANKA.

Fax: (+94 8) 389 106
E-mail: director@4wch.com

Instructions to Contributors

Manuscripts in English should be typed on sturdy typing paper of A4 size (28 ※ 21cm).
Typing should be double-spaced with 2.5 cm margins on all sides. Words should not be broken
at the end of a line. Full papers should not exceed 12 printed pages, and short communications
should not exceed 2 printed pages, Longer papers may be printed, on condition that the author
pays the excess page charges in full. Each of the following divisions should be begun on a
separate page: cover page, title page, abstract, main text, references, each table, figure legends,
each figure. The cover page should show the title, name of author and addresses to whom com-
munications should be directed with his or her telephone and fax numbers, and E-mail address,
the date of submission, the number of pages of main text, the number of figure legends, number
of figures, and number of tables.

The title page should show the title, the name(s) of the author(s), and their affiliation(s) and
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119

AoOMWarsiTo. oO} nologies?

7
ーー

- -
の

CURRENT HERPETOLOGY

MANAGING EDITOR
Masafumi MATSUI

Graduate School of Human and Environmental Studies, Kyoto University, Yoshida
Nihonmatsu-cho, Sakyo-ku, Kyoto 606-8501 Japan
(fumi@zoo.zool.kyoto-u.ac.jp)

ASSOCIATE EDITORS

Kraig ADLER, Department of Neurobiology and Behavior, Cornell University, Seeley G. Mudd
Hall, Ithaca, New York 14853-2702, USA (kka4@cornell.edu)

Aaron M. BAUER, Department of Biology, Villanova University, 800 Lancaster Avenue, Vil-
lanova, PA 19085 USA (aaron.bauer@villanova.edu)

Ilya S. DAREVSKY, Zoological Institute, Russian Academy of Sciences, St.Petersburg 199034
RUSSIA (Darevsky@herpet.zin.ras.spb.ru)

twics.com)

Ivan INEICH, Laboratoire des Reptiles et Amphibiens, Museum National d’ Histoire Naturelle,
25 rue Cuvier 75005 Paris, FRANCE (ineich@cimrs1.mnhn.fr)

Ulrich JOGER, Hessisches Landesmuseum Darmstadt, Zoologische Abteilung, Friedensplatz 1,
D-64283 Darmstadt, GERMANY (u.joger@hlmd.tu-darmstadt.de)

Tamotsu KUSANO, Department of Biological Science, Graduate School of Science, Tokyo
Metropolitan University, Minami-ohsawa, Hachioji, Tokyo 192-0397 JAPAN (tamo@
comp.metro-u.ac.jp)

Colin McCARTHY, Department of Zoology, The Natural History Museum, Cromwell road,
London SW7 5BD, UK (cjm@nhm.ac.uk)

Hidetoshi OTA, Tropical Biosphere Research Center, University of the Ryukyus, Nishihara,
Okinawa, 903-0213 JAPAN (ota@sci.u-ryukyu.ac.jp)

Michihisa TORIBA, Japan snake Institute, Yabuzuka-honmachi, Nitta-gun, Gunma 379-2300
JAPAN (snake-a@sunfield.ne.jp)

Home Page of THE HERPETOLOGICAL SOCIETY OF JAPAN
http://zoo.zool.kyoto-u.ac.jp/~herp/

DATE OF PUBLICATION
Current Herpetology, Vol. 19, No. 1,
was mailed 15 July 2000.

ーーーー テ ーー テー

まい す 、 コ ー
_a |

iii

8 1387

CONTENTS

Originals

Phylogenetic position of Draco fimbriatus, with a molecular perspective on the
historical biogeography of the genus Draco (Reptilia: Agamidae) |
CD と Masanao Honda, Hidetoshi Ota, Showichi Sengoku, and Tsutomu Hikida::::-- 43

Mabuya cumingi (Reptilia: Scincidae): An addition to the herpetofauna of Lanyu Island,
Taiwan ee SREY ac AE OCH HOPE coin ote FIONN eh ORE con orm copa の Hidetoshi Ota and Wen-San Huang Stal cate 57

Sperm morphology of some Indian forgs as revealed by SEM
ao se/ る So の PPS、 niece FMS mae iedehet Mandate: drakaveee heucrelehete co eh ciamene nualelop sin Mitsuru Kuramoto and S. Hareesh Joshy: wets 63

On the monophyly of the agamid genus Gonocephalus Kaup, 1825 (Reptilia: Squamata):
A chromosomal perspective:::::: Cheong-Hoong Diong, May-Hon Low,
Ene-Choo Tan, Hoi-Sen Yong, Tsutomu Hikida, and Hidetoshi Ota:::::- a

Varidity of back-calculation methods of body size from phalangeal bones: An assessment
using data for Rana japonica:::::: Junsuke Marunouchi, Tamotsu Kusano, and

Hiroaki Ueda:::::: 81 ;

Relationship between brightness or size and the presence of barnacles on the carapace

of the hawksbill turtles (Eretmochelys 777 の 77c272) で … ド mm Mari Kobayashi:::::-
Review
Literature survey on predators of snakes in Japan -:::::::: Koji Tanaka and Akira Mori:::::: 111
Titles of Papers Presented at Annual Meeting oo 113
CGY Tet Ree eer rere ener roc as ry ee bk,

FUTURE MEETING
Niigata University, Niigata, Japan, November 2001
(Kunio Sekiya, Chair)

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